>From cthummel@XXXX Thu Jun 20  12:27:46  1996
Date: 20 Jun 1996  10:26:13  U
From: 'CThummel' <cthummel@XXXX>
Subject: pCaSpeR vectors
To: 'Madeline Crosby' <crosby@XXXX>
X-Mailer: Mail*Link SMTP-QM 3.0.2
Content-Length: 905
  REGARDING           pCaSpeR vectors
Dear Lynn,
You may remember that you contacted me a year ago regarding the sequences of P
element vectors that we made, together with Vince Pirrotta.  We have been
working on those sequences (very slowly!) and finally have some accession
numbers for you.  We also have the maps of these vectors (and accession
numbers) on a WWW page
(http://www-hhmi.genetics.utah.edu/thummel/pelement.html) in case you are
interested.  The numbers are as follows:
pCaSpeR  EMBL 81644
pCaSpeR2  GenBank U59054
pCaSpeR3  GenBank U59055
pCaSpeR4  EMBL 81645
pCaSpeR-hs  GenBank U59056
pCaSpeR-hs/act GenBank U60735
pCaSpeR hs43 lacZ  EMBL 81643
I hope that this helps!  We will let you know if we find the energy to
assemble sequences of any lacZ vectors.  I think it would be nice to have
pCaSpeR-AUG-bgal, but I can't get too excited about it right now!  
Best wishes,
Carl Thummel
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>From crosby Thu Jun 20  15:35:54  1996
Date: Thu, 20 Jun 96  15:35:49  EDT
From: crosby (Madeline Crosby)
To: cthummel@XXXX
Subject: Re: pCaSpeR vectors
Cc: crosby
Content-Type: X-sun-attachment
Content-Length: 29725
Carl,
Many thanks for remembering my request!  Your timing could not be
better: FlyBase is planning on publishing maps of commonly used
vectors, enhancer traps, etc., in an upcoming FlyBase issue of DIS.
I am not familiar with the details on permission from authors, etc.,
but I hope we can include these vectors.
In regard to the lacZ vectors: I already have essentially complete
compiled sequences for these.  (Complete in the case of CaSpeR-betagal;
a few unknown nucleotides in the case of CaSpeR-AUG-betagal.)  Would you 
like to look them over and see what you think (attached below)?  I know 
that Bill would like to work out a system of joint submissions to GenBank; 
do you think this might be a reasonable option for these two vectors?
Another question: below is a comment I have included in my compilation
of the white sequences in the CaSpeR vectors.  Do you know anything
about these discrepancies?
' Sequence discrepancies (+1 bp at 2176; c to t at undetermined position) 
   in sequence entry listed versus white (sequence entry X02974 
   [Genbank / EMBL] | 10873 | 22 Feb 1993). FlyBase-compiled sequence is 
   based upon the latter and thus differs from sequence in entry X81644.'
Thanks again.
Sincerely,
Lynn
Lynn Crosby
FlyBase
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>From cthummel@XXXX Fri Jun 21  17:39:38  1996
Date: 21 Jun 1996  15:37:25  U
From: 'CThummel' <cthummel@XXXX>
Subject: Re: pCaSpeR vectors
To: 'Madeline Crosby' <crosby@XXXX>
X-Mailer: Mail*Link SMTP-QM 3.0.2
Content-Type: x-binhex4
X-Attachments: 'Casper sequences' (type: binhex4)
Content-Length: 34414
                      RE>>pCaSpeR vectors                          6/21/96
Hi Lynn,
My, you have been busy!  Needless to say, I did not expect that anyone would
put the time and effort necessary into the compilation of these sequences,
much less someone who had not actually made the clones themselves!  There are
a lot of little cloning tricks in there that are tricky to figure out - even
for me!!  Anyhow, the basic sequences you sent look OK.  There are, however, a
few problems.  First, the length of both vectors is off by several kb - they
should each be about 12 kb total.  The sequence you are missing is in the
Casper vector.  You should replace your Casper sequences in any case, since
there have been several recent small changes that I learned about from Vince
Pirrotta.  I have enclosed the most recent Casper sequence, along with a
modified version of your Casper-bgal vector.  You had a run of 4 Cs downstream
from the polylinker at the 5' end of lacZ - this should have been 3 Cs.  The
sequence was correct in your Casper-AUG-bgal sequence (probably because there
you could see that the Adh AUG needed to be in frame with lacZ).  Otherwise,
the only parts I could not confirm were the Casper-SV40, SV40-lacZ junctions
in both sequences, and Casper-Adh-lacZ junction in Casper-AUG-bgal.  I think
it would be safest to sequence through these junctions.  Did you actually do
that, or is your sequence just a best guess based on the literature?  Some of
these junctions are impossible to predict without sequencing.  We would be
happy to do the sequencing, if you don't mind waiting a bit.  Please let me
know what you think!  Thanks for all your help getting this together.
Best wishes,
Carl
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>From crosby Fri Jun 21  18:12:49  1996
Date: Fri, 21 Jun 96  18:12:43  EDT
From: crosby (Madeline Crosby)
To: cthummel@XXXX
Subject: Re: pCaSpeR vectors
Cc: crosby
Content-Length: 2372
Carl,
We're making progress!
>First, the length of both vectors is off by several kb - they
>should each be about 12 kb total.
The CaSpeR vector sequences are not included in these -- they
are the transposon-only versions.  A slightly cumbersome but
occasionally useful convention we have adopted is to 
represent everything (except construction intermediates) in
two ways: both the plasmid (or cosmid) form and the transposon 
form.  I hesitated to send you the plasmid form because it is
less obvious which is the best orientation and what is the
best starting point.
The sizes I have currently listed are:
pP{CaSpeR-betagal}      11.978kb
P{CaSpeR-betagal}        9.218kb
pP{CaSpeR-AUG-betagal} ~12.126kb
P{CaSpeR-AUG-betagal    ~9.366kb
I know the ~ with all significant digits looks a bit strange, but
the size is derived automatically by adding up component segments.
> ...is your sequence just a best guess based on the literature?
Yes.
>Some of these junctions are impossible to predict without sequencing.
Very true!  Initially, I was trying to give some sort of confidence
rating on the compiled sequences, but that really isn't practical.
For things that are REALLY dubious, I include disclaimers in the
comments sections.  It might be a good idea to include some sort of
generic disclaimer, along with a request for corrections when errors
are discovered.
>We would be happy to do the sequencing, if you don't mind waiting a bit.
That would be terrific!  Presumably these small stretches can be
done relatively quickly.
>You should replace your Casper sequences in any case, since
>there have been several recent small changes that I learned about from Vince
>Pirrotta.
Is Vince going to correct his GenBank/EMBL entries?  Large stretches of 
sequence like this I have been pulling from GenBank; we store only the 
compiled sequences and these are re-compiled periodically to reflect any 
changes that have been submitted to GenBank.  With only a few exceptions, 
the only original sequence that we store is that of junctions.
Thanks for all the input.  Let me know how the sequencing goes.
--Lynn
P.S. I get the distinct impression that you haven't looked at
TransposonSearch on FlyBase.  It can only be accessed through the
Web, but if you can manage to take a look at what we've been doing,
I would really appreciate your feedback.
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>From cthummel@XXXX Mon Jun 24  12:00:49  1996
Date: 24 Jun 1996  09:59:42  U
From: 'CThummel' <cthummel@XXXX>
Subject: Re: pCaSpeR vectors
To: 'Madeline Crosby' <crosby@XXXX>
X-Mailer: Mail*Link SMTP-QM 3.0.2
Content-Length: 4066
                      RE>>pCaSpeR vectors                          6/24/96
Hi Lynn,
Thanks for your response.  I have browsed through Transposon search and am
very impressed.  I must confess that although I use FlyBase several times each
day, I have not tried that handy little window!  Why don't you go ahead and
submit your sequences with a disclaimer.  We will then go ahead and sequence
the junctions that I mentioned to you earlier.  Although it should be quick,
it won't be.  We have a bunch of other stuff going on and will fit these
sequencing reactions in as we can.  I will let you know once we have assembled
a complete corrected sequence for both Casper-bgal and Casper-AUG-bgal. 
Thanks again,
Carl
P.S. Vince will be correcting his EMBL submissions, so your re-compilation of
the sequence should incorporate his changes.  Keep up the terrific work!
----- End Included Message -----
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>From cthummel@XXXX Mon Jul 29  16:50:12  1996
Date: 29 Jul 1996  14:51:15  U
From: 'CThummel' <cthummel@XXXX>
Subject: Casper-bgal sequences
To: 'Lynn Crosby' <crosby@XXXX>
X-Mailer: Mail*Link SMTP-QM 3.0.2
Content-Length: 19532
  REGARDING           Casper-bgal sequences
Hi Lynn,
We have finished sequencing the junctions on pCasper-bgal and
pCasper-AUG-bgal.  We did the lacZ-SV40 junctions on both vectors, the
SV40-Casper junctions on both vectors, and the Casper-AUG-lacZ junctions on
pCasper-AUG-bgal.  As expected, there were small insertions and/or deletions
to make in these regions.  I have enclosed the modified sequences.  I hope
that these help!  Please send me the accession numbers once you submit them to
GenBank.  Thanks again for your help in getting these sequences together!
Best wishes,
Carl
<<<<<< Attached TEXT file named 'cas-aug-bgal' follows >>>>>>
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tttgtagaggttttacttgctttaaaaaacctcccacacctccccctgaacctgaaacataaaatgaatgcaattgttgt
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cctgcagcccaagctttgcgtactcgcaaattattaaaaataaaactttaaaaataatttcgtctaattaatattatgag
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aatactattcctttcactcgcacttattgcaagcatacgttaagtggatgtctcttgccgacgggaccaccttatgttat
ttcatcatg
<<<<<< Attached TEXT file named 'cas-bgal' follows >>>>>>
catgatgaaataacataaggtggtcccgtcggcaagagacatccacttaacgtatgcttgcaataagtgcgagtgaaagg
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ggaaagctggctggagtgcgatcttcctgaggccgatactgtcgtcgtcccctcaaactggcagatgcacggttacgatg
cgcccatctacaccaacgtaacctatcccattacggtcaatccgccgtttgttcccacggagaatccgacgggttgttac
tcgctcacatttaatgttgatgaaagctggctacaggaaggccagacgcgaattatttttgatggcgttaactcggcgtt
tcatctgtggtgcaacgggcgctgggtcggttacggccaggacagtcgtttgccgtctgaatttgacctgagcgcatttt
tacgcgccggagaaaaccgcctcgcggtgatggtgctgcgttggagtgacggcagttatctggaagatcaggatatgtgg
cggatgagcggcattttccgtgacgtctcgttgctgcataaaccgactacacaaatcagcgatttccatgttgccactcg
ctttaatgatgatttcagccgcgctgtactggaggctgaagttcagatgtgcggcgagttgcgtgactacctacgggtaa
cagtttctttatggcagggtgaaacgcaggtcgccagcggcaccgcgcctttcggcggtgaaattatcgatgagcgtggt
ggttatgccgatcgcgtcacactacgtctgaacgtcgaaaacccgaaactgtggagcgccgaaatcccgaatctctatcg
tgcggtggttgaactgcacaccgccgacggcacgctgattgaagcagaagcctgcgatgtcggtttccgcgaggtgcgga
ttgaaaatggtctgctgctgctgaacggcaagccgttgctgattcgaggcgttaaccgtcacgagcatcatcctctgcat
ggtcaggtcatggatgagcagacgatggtgcaggatatcctgctgatgaagcagaacaactttaacgccgtgcgctgttc
gcattatccgaaccatccgctgtggtacacgctgtgcgaccgctacggcctgtatgtggtggatgaagccaatattgaaa
cccacggcatggtgccaatgaatcgtctgaccgatgatccgcgctggctaccggcgatgagcgaacgcgtaacgcgaatg
gtgcagcgcgatcgtaatcacccgagtgtgatcatctggtcgctggggaatgaatcaggccacggcgctaatcacgacgc
gctgtatcgctggatcaaatctgtcgatccttcccgcccggtgcagtatgaaggcggcggagccgacaccacggccaccg
atattatttgcccgatgtacgcgcgcgtggatgaagaccagcccttcccggctgtgccgaaatggtccatcaaaaaatgg
ctttcgctacctggagagacgcgcccgctgatcctttgcgaatacgcccacgcgatgggtaacagtcttggcggtttcgc
taaatactggcaggcgtttcgtcagtatccccgtttacagggcggcttcgtctgggactgggtggatcagtcgctgatta
aatatgatgaaaacggcaacccgtggtcggcttacggcggtgattttggcgatacgccgaacgatcgccagttctgtatg
aacggtctggtctttgccgaccgcacgccgcatccagcgctgacggaagcaaaacaccagcagcagtttttccagttccg
tttatccgggcaaaccatcgaagtgaccagcgaatacctgttccgtcatagcgataacgagctcctgcactggatggtgg
cgctggatggtaagccgctggcaagcggtgaagtgcctctggatgtcgctccacaaggtaaacagttgattgaactgcct
gaactaccgcagccggagagcgccgggcaactctggctcacagtacgcgtagtgcaaccgaacgcgaccgcatggtcaga
agccgggcacatcagcgcctggcagcagtggcgtctggcggaaaacctcagtgtgacgctccccgccgcgtcccacgcca
tcccgcatctgaccaccagcgaaatggatttttgcatcgagctgggtaataagcgttggcaatttaaccgccagtcaggc
tttctttcacagatgtggattggcgataaaaaacaactgctgacgccgctgcgcgatcagttcacccgtgcaccgctgga
taacgacattggcgtaagtgaagcgacccgcattgaccctaacgcctgggtcgaacgctggaaggcggcgggccattacc
aggccgaagcagcgttgttgcagtgcacggcagatacacttgctgatgcggtgctgattacgaccgctcacgcgtggcag
catcaggggaaaaccttatttatcagccggaaaacctaccggattgatggtagtggtcaaatggcgattaccgttgatgt
tgaagtggcgagcgatacaccgcatccggcgcggattggcctgaactgccagctggcgcaggtagcagagcgggtaaact
ggctcggattagggccgcaagaaaactatcccgaccgccttactgccgcctgttttgaccgctgggatctgccattgtca
gacatgtataccccgtacgtcttcccgagcgaaaacggtctgcgctgcgggacgcgcgaattgaattatggcccacacca
gtggcgcggcgacttccagttcaacatcagccgctacagtcaacagcaactgatggaaaccagccatcgccatctgctgc
acgcggaagaaggcacatggctgaatatcgacggtttccatatggggattggtggcgacgactcctggagcccgtcagta
tcggcggaccagctgagcgccggtcgctaccattaccagttggtctggtgtcaaaaataataataaccgggcaggccatg
tctgcccgtatttcgcgtaaggaaatccattatgtactatttaaaaaacacaaacttttggatgttcggtttattctttt
tcttttacttttttatcatgggagcctacttcccgtttttcccgatttggctacatgacatcaaccatatcagcaaaagt
gatacgggtattatttttgccgctatttctctgttctcgctattattccaaccgctgtttggtctgctttctgacaaact
cggcctcgactctagctagaggatctttgtgaaggaaccttacttctgtggtgtgacataattggacaaactacctacag
agatttaaagctctaaggtaaatataaaatttttaagtgtataatgtgttaaactactgattctaattgtttgtgtattt
tagattccaacctatggaactgatgaatgggagcagtggtggaatgcctttaatgaggaaaacctgttttgctcagaaga
aatgccatctagtgatgatgaggctactgctgactctcaacattctactcctccaaaaaagaagagaaaggtagaagacc
ccaaggactttccttcagaattgctaagttttttgagtcatgctgtgtttagtaatagaactcttgcttgctttgctatt
tacaccacaaaggaaaaagctgcactgctatacaagaaaattatggaaaaatattctgtaacctttataagtaggcataa
cagttataatcataacatactgttttttcttactccacacaggcatagagtgtctgctattaataactatgctcaaaaat
tgtgtacctttagctttttaatttgtaaaggggttaataaggaatatttgatgtatagtgccttgactagagatcataat
cagccataccacatttgtagaggttttacttgctttaaaaaacctcccacacctccccctgaacctgaaacataaaatga
atgcaattgttgttgttaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaat
aaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctggatcgggcgagc
tcgaattctagtatgtatgtaagttaataaaacccttttttggagaatgtagatttaaaaaaacatattttttttttatt
ttttactgcactggacatcattgaacttatctgatcagttttaaatttacttcgatccaagggtatttgaagtaccaggt
tctttcgattacctctcactcaaaatgacattccactcaaagtcagcgctgtttgcctccttctctgtccacagaaatat
cgccgtctctttcgccgctgcgtccgctatctctttcgccaccgtttgtagcgttacctagcgtcaatgtccgccttcag
ttgcactttgtcagcggtttcgtgacgaagctccaagcggtttacgccatcaattaaacacaaagtgctgtgccaaaact
cctctcgcttcttatttttgtttgttttttgagtgattggggtggtgattggttttgggtgggtaagcaggggaaagtgt
gaaaaatcccggcaatgggccaagaggatcaggagctattaattcgcggaggcagcaaacacccatctgccgagcatctg
aacaatgtgagtagtacatgtgcatacatcttaagttcacttgatctataggaactgcgattgcaacatcaaattgtctg
cggcgtgagaactgcgacccacaaaaatcccaaaccgcaatcgcacaaacaaatagtgacacgaaacagattattctggt
agctgtgctcgctatataagacaatttttaagatcatatcatgatcaagacatctaaaggcattcattttcgactacatt
cttttttacaaaaaatataacaaccagatattttaagctgatcctagatgcacaaaaaataaataaaagtataaacctac
ttcgtaggatacttcgttttgttcggggttagatgagcataacgcttgtagttgatatttgagatcccctatcattgcag
ggtgacagcggacgcttcgcagagctgcattaaccagggcttcgggcaggccaaaaactacggcacgctcctgccaccca
gtccgccggaggactccggttcagggagcggccaactagccgagaacctcacctatgcctggcacaatatggacatcttt
ggggcggtcaatcagccgggctccggatggcggcagctggtcaaccggacacgcggactattctgcaacgagcgacacat
accggcgcccaggaaacatttgctcaagaacggtgagtttctattcgcagtcggctgatctgtgtgaaatcttaataaag
ggtccaattaccaatttgaaactcagtttgcggcgtggcctatccgggcgaacttttggccgtgatgggcagttccggtg
ccggaaagacgaccctgctgaatgcccttgcctttcgatcgccgcagggcatccaagtatcgccatccgggatgcgactg
ctcaatggccaacctgtggacgccaaggagatgcaggccaggtgcgcctatgtccagcaggatgacctctttatcggctc
cctaacggccagggaacacctgattttccaggccatggtgcggatgccacgacatctgacctatcggcagcgagtggccc
gcgtggatcaggtgatccaggagctttcgctcagcaaatgtcagcacacgatcatcggtgtgcccggcagggtgaaaggt
ctgtccggcggagaaaggaagcgtctggcattcgcctccgaggcactaaccgatccgccgcttctgatctgcgatgagcc
cacctccggactggactcatttaccgcccacagcgtcgtccaggtgctgaagaagctgtcgcagaagggcaagaccgtca
tcctgaccattcatcagccgtcttccgagctgtttgagctctttgacaagatccttctgatggccgagggcagggtagct
ttcttgggcactcccagcgaagccgtcgacttcttttcctagtgagttcgatgtgtttattaagggtatctagcattaca
ttacatctcaactcctatccagcgtgggtgcccagtgtcctaccaactacaatccggcggacttttacgtacaggtgttg
gccgttgtgcccggacgggagatcgagtcccgtgatcggatcgccaagatatgcgacaattttgctattagcaaagtagc
ccgggatatggagcagttgttggccaccaaaaatttggagaagccactggagcagccggagaatgggtacacctacaagg
ccacctggttcatgcagttccgggcggtcctgtggcgatcctggctgtcggtgctcaaggaaccactcctcgtaaaagtg
cgacttattcagacaacggtgagtggttccagtggaaacaaatgatataacgcttacaattcttggaaacaaattcgcta
gattttagttagaattgcctgattccacacccttcttagtttttttcaatgagatgtatagtttatagttttgcagaaaa
taaataaatttcatttaactcgcgaacatgttgaagatatgaatattaatgagatgcgagtaacattttaatttgcagat
ggttgccatcttgattggcctcatctttttgggccaacaactcacgcaagtgggcgtgatgaatatcaacggagccatct
tcctcttcctgaccaacatgacctttcaaaacgtctttgccacgataaatgtaagtcttgtttagaatacatttgcatat
taataatttactaactttctaatgaatcgattcgatttaggtgttcacctcagagctgccagtttttatgagggaggccc
gaagtcgactttatcgctgtgacacatactttctgggcaaaacgattgccgaattaccgctttttctcacagtgccactg
gtcttcacggcgattgcctatccgatgatcggactgcgggccggagtgctgcacttcttcaactgcctggcgctggtcac
tctggtggccaatgtgtcaacgtccttcggatatctaatatcctgcgccagctcctcgacctcgatggcgctgtctgtgg
gtccgccggttatcataccattcctgctctttggcggcttcttcttgaactcgggctcggtgccagtatacctcaaatgg
ttgtcgtacctctcatggttccgttacgccaacgagggtctgctgattaaccaatgggcggacgtggagccgggcgaaat
tagctgcacatcgtcgaacaccacgtgccccagttcgggcaaggtcatcctggagacgcttaacttctccgccgccgatc
tgccgctggactacgtgggtctggccattctcatcgtgagcttccgggtgctcgcatatctggctctaagacttcgggcc
cgacgcaaggagtagccgacatatatccgaaataactgcttgtttttttttttaccattattaccatcgtgtttactgtt
tattgccccctcaaaaagctaatgtaattatatttgtgccaataaaaacaagatatgacctatagaatacaagtatttcc
ccttcgaacatccccacaagtagactttggatttgtcttctaaccaaaagacttacacacctgcataccttacatcaaaa
actcgtttatcgctacataaaacaccgggatatattttttatatacatacttttcaaatcgcgcgccctcttcataattc
acctccaccacaccacgtttcgtagttgctctttcgctgtctcccacccgctctccgcaacacattcaccttttgttcga
cgaccttggagcgactgtcgttagttccgcgcgattcggttcgctcaaatggttccgagtggttcatttcgtctcaatag
aaattagtaataaatatttgtatgtacaatttatttgctccaatatatttgtatatatttccctcacagctatatttatt
ctaatttaatattatgactttttaaggtaattttttgtgacctgttcggagtgattagcgttacaatttgaactgaaagt
gacatccagtgtttgttccttgtgtagatgcatctcaaaaaaatggtgggcataatagtgttgtttatatatatcaaaaa
taacaactataataataagaatacatttaatttagaaaatgcttggatttcactggaactagaattaattcggctgctgc
tctaaacgacgcatttcgtactccaaagtacgaattttttccctcaagctcttattttcattaaacaatgaacaggacct
aacgcacagtcacgttattgtttacataaatgattttttttactattcaaacttactctgtttgtgtactcccactggta
tagccttcttttatcttttctggttcaggctctatcactttactaggtacggcatctgcgttgagtcgcctccttttaaa
tgtctgaccttttgcaggtgcagccttccactgcgaatcattaaagtgggtatcacaaatttgggagttttcaccaaggc
tgcacccaaggctctgctcccacaattttctcttaatagcacacttcggcacgtgaattaattttactccagtcacagct
ttgcagcaaaatttgcaatatttcatttttttttattccacgtaagggttaatgttttcaaaaaaaaattcgtccgcaca
caacctttcctctcaacaagcaaacgtgcactgaatttaagtgtatacttcggtaagcttcggctatcgacgggaccacc
ttatgttatttcatcatg
----- End Included Message -----
----- Begin Included Message -----
>From crosby Sun Aug  4  18:43:01  1996
Date: Sun, 4 Aug 96  18:42:59  EDT
From: crosby (Madeline Crosby)
To: cthummel@XXXX
Subject: Re: Casper-bgal sequences
Cc: crosby
Content-Length: 1134
Carl,
Thanks! (Sorry to be a bit slow to reply -- I've been on vacation.)
These corrections should be incorporated into the FlyBase-compiled
sequences within the next week or two.
Two questions: 
1) In pCaSpeR-AUG-Bgal your corrected junction between SV40 and CaSpeR
   includes an EcoRI site.  If this is correct, EcoRI can not
   be used as a cloning site.  Are there, indeed, two EcoRI sites??
2) Is the junction between lacZ and SV40 in pCaSpeR-Bgal identical to 
   that in CaSpeR-hs43-lacZ?  
For the GenBank submissions I think I would prefer not to use the 
standard FlyBase format (P transposons always presented P5' to P3' and
vectors broken within the polylinker). I am inclined to prepare them in 
the orientation you have your sequences (with the lacZ 5' to 3'), 
starting with the P sequences and appending the vector sequences.  So, 
unless I hear any objections....
Who should be included in the Thummel lab citation?  I assume we
should cite the Thummel et al. 1988 Gene paper, and for CaSpeR-Bgal, 
the DIN/DIS correction.  Any others?
I'm really pleased that we were able to pull this together!
--Lynn
----- End Included Message -----
----- Begin Included Message -----
Date: 5 Aug 1996  11:18:51  U
From: 'CThummel' <cthummel@XXXX>
Subject: Re: Casper-bgal sequences
To: 'Madeline Crosby' <crosby@XXXX>
X-Mailer: Mail*Link SMTP-QM 3.0.2
Content-Length: 3289
                      RE>>Casper-bgal sequences                    8/5/96
Thanks Lynn - I am glad to hear that you take vacations!  I would imagine that
staring at sequences gets to be a little tedious after awhile!  Sorry about
the error in the SV40 CaSpeR junction of pCaSpeR-AUG-Bgal.  I was just
focusing on the junction sequences.  There is a base change in the CaSpeR
sequence, immediately downstream from the junction that inactivates the EcoRI
site.  The sequence should read: ...GAATTGGTCGACC...  Unfortunately, I found a
few other single base changes further downstream in CaSpeR, but I do not want
to get involved in proofing all 7.8 kb!!  What a pain.
The lacZ/SV40 junctions are different between CaSpeR-Bgal and
CaSpeR-hs43-lacZ.  Vince Pirrotta independently constructed the latter vector,
using pCaSpeR4.  That accounts for the 6 bp difference between the two
junction sequences.  
I agree with you, that it would be best to have the vector sequences present
in the GenBank submission, and have the lacZ sequences ordered 5' to 3'.  For
citations, I think that the Gene paper is sufficient (and DIN/DIS correction
for pCaSpeR-Bgal).  Thanks for all your help!  I am so happy that this got
done - I continue to get requests for these sequences.  Please let me know
when you have accession numbers,
Carl
----- End Included Message -----
----- Begin Included Message -----
>From crosby Wed Aug  7  10:56:31  1996
Date: Wed, 7 Aug 96  10:56:29  EDT
From: crosby (Madeline Crosby)
To: cthummel@XXXX
Subject: Casper-AUG-bgal sequences
Cc: crosby
Content-Length: 515
Carl,
Sorry to be harrassing you, again, but this is a little disturbing: 
> Unfortunately, I found a
>few other single base changes further downstream in CaSpeR, but I do not want
>to get involved in proofing all 7.8 kb!!
There could easily be errors within the polylinker sequence, which is
only another 20-25bp.  (Many necessary fill-ins, etc., performed during
constructions never get mentioned.) These I would like to know about.  
The sequence beyond that should be okay (P3' published sequence).
--Lynn
----- End Included Message -----
----- Begin Included Message -----
>From cthummel@XXXX Wed Aug  7  12:10:35  1996
Date: 7 Aug 1996  10:10:20  U
From: 'CThummel' <cthummel@XXXX>
Subject: Re: Casper-AUG-bgal sequence
To: 'Madeline