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ID   PHS85      preliminary; circular DNA; SYN; 9600 BP.
XX
AC   M32616; ATCC37842;
XX
DT   15-AUG-1990 (Rel. 5, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Drosophila/E.coli plasmid vector pHS85 - incomplete, hsp82-neo.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-3727
RC   pHS1 from pMK16 & pKM109/90, Tn5 neo gene
RC   pHS2 from pHS1
RC   pHS7 from pBR322 & linker
RC   pHS8 from pHS7 & pHS2
RC   pHS9 from p82R & pBR322
RC   pHS10 from pHS8 & pHS9
RC   pHS11 from pBR322 & pHS10
RC   pHS13 from pHS11
RC   pHS16 from pHS13 & p906.2
RC   pHS17 from pHS16 & linker
RC   pHS22 from pAP-2 & pHS, hsp82-neo-polyA fragment
RC   pHS24 from pHS17 & pAP-2
RC   pHS82 from pBluescript KS+ & pPLdelta-1
RC   pHS83 from pHS82
RC   pHS84 from pHS83 & linker
RC   pHS85 from pHS84 & pHS, hsp82-neo-polyA fragment
RC   pHS86 from pBluescript KS+ & pHS, hsp82-neo-polyA fragment
RA   Sass H.;
RT   "P-transposable vectors expressing a constitutive and
RT   thermoinducible hsp82-neo fusion gene for Drosophila germline
RT   transformation tissue-culture transfection";
RL   Gene 89:179-186(1990).
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RN   [2]
RC   pHS85
RA   Sass H.;
RT   ;
RL   Submitted (06-MAR-1990) to GenBank on computer-readable media by:
RL   Sass H., .
XX
CC   P-transposable (integrating) shuttle vector. Contains hsp82-neo
CC   fusion gene which encodes the first 20 aa of D. pseudoobscura hsp82,
CC   5 codons in a linker, fused to the fifth codon of bacterial neomycin
CC   phosphotransferase from Tn5, and followed by polyadenylation signals
CC   from D. melanogaster hsp82. Also contains an 868 bp hsp82 promoter
CC   regulating the hsp82-neo fusion gene and a 102 bp multiple cloning
CC   site between the polyadenylation signal and the inverted repeats of
CC   P-element ppi25.1.
CC   Expression is constitutive, but can be increased with heat.
CC   Selection of transgenic Drosophila by G418 has been done with
CC   embryos, larvae, and adults.
CC   Can be used in Drosophila spp. cell culture transfection assays, and
CC   may function in other animal and plant systems
CC   because the hsp82 promoter is highly conserved.
CC   Deposited by: H.G.Sass
CC   Growth media: LB plus ampicillin (ATCC medium number 1227) 37C
CC   NM (pHS85)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (Drosophila)
CC   CP ()
CC   FN (transfection)(expression thermoinducible)
CC   SE ()
CC   PA (P element)(D.pseudoobscura)(D.melanogaster)(pHS85)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. ppi25.1, P element
FT                   -> pPLdelta-1
FT                   1. pBluescript KS+ SacI-KpnI 102bp 658..760, MCS
FT                   2. pPLdelta-1 remove small KpnI-SacI
FT                   -> pHS82
FT                   1. pHS82 KpnI, MCS
FT                   T4 DNA polymerase
FT                   -> pHS83
FT                   1. pHS83 SacI, MCS
FT                   T4 DNA polymerase
FT                   KpnI linker 8bp nggtaccn
FT                   -> pHS84 [unique KpnI in 3' MCS]
FT                   1. pHS17 ClaI-BamHI 800bp, no hsp82 5' to -865
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pHS[neo-4.2.2.KpnI(3')/BamHI(-865)] [unique BamHI]
FT                   1. pHS84 remove BglII-KpnI, 5' MCS/5870bp
FT                   2. pHS[neo-4.2.2.KpnI(3')/BamHI(-865)]
FT                   BamHI-KpnI 3730bp, hsp82-neo-polyA fragment
FT                   -> pHS85 9600bp"
FT   misc_feature    6..868
FT                   /note="D. pseudoobscura hsp82 gene non-coding
FT                   5' flank"
FT   misc_feature    869..1004
FT                   /note="D. pseudoobscura heat-shock protein 82 (hsp82)
FT                   exon 1"
FT   intron          1005..2067
FT                   /note="D. pseudoobscura hsp82 gene intron A"
FT   CDS             2068..2925
FT                   /note="GEN D.pseudoobscura heat-shock protein 82/
FT                   neomycin phosphotransferase fusion protein
FT                   (hsp82-neo)"
FT   misc_feature    2068..2126
FT                   /note="D. pseudoobscura heat-shock protein 82 (hsp82)
FT                   truncated exon 2"
FT   misc_binding    2127..2142
FT                   /note="MCS linker"
FT   CDS             2143..3269
FT                   /note="ANT E. coli neomycin phosphotransferase gene
FT                   (NPT), neomycin resistance gene (neo)"
FT   polyA_signal    3270..3610
FT                   /note="PLA D. melanogaster hsp82 gene non-coding
FT                   3' flank"
FT   misc_binding    3620..3726
FT                   /note="MCS"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
XX
SQ   Sequence 3727 BP; 950 A; 866 C; 882 G; 1029 T; 0 other;
     ggatccgatg gatttttacc atattattat tatttctagc cacgttgcaa ctctatgtca
     gtaccggaaa tagcagccct ggagtctctt agcctctaga aacggctaga acattctacg
     cttgtggttg gttttcattg aaagcaggcg tcttttatat actttacggt atatagctac
     atgtatataa tggtatactt catcaatatc atcaatctat gaattttaat ttttaagagt
     acatatataa attaacatgg gggatatagt tctcaatacc caagtatttg aattttccat
     ctctcatcgg gggtaattca tgaaccggtt ccagccgaaa aatgaacgaa attcatgaga
     gattattttt tcgggattgc ttgccaatac atttcggaaa aacaaaatgt actacatttt
     tgtcatctca gggtgctcca attaattatg aatgctacga cactacaaag cagcttggaa
     atccgaattt taacaataat taaaggaaat agggtatagc gtatataggg tatcatagct
     gaaacgggta taccaacaat aatgacgcag cacttacgtt tcactccgta ctcacttacg
     atttatgctt ataatttttg ttcacctctt ttacttaaac ctcactttaa aaacaatcaa
     ataaatggga gtatttatgt atatttctaa gattacggcg gtattgttct gctgtctgcg
     gtcacactgg ttttcagcct cggtgcaact ctgtttcagt accggaaata gcagccctgg
     attctcgtag cctctagaaa cgtctagaaa attctacgct tggggttggt ttgctataaa
     agcaggcggg ccgactgttg ccggctcgag tcttgaaaaa tttttgtcca gtgaaggtgc
     gtttgcttag agcgcagtgc aacaaagtga atttattcta cacaaatcga agtgaaaata
     tatatatatt tttatctctg ctgttaaatt aaaacacata caaggtaagc gttaacaatg
     aaagtgcatt tatttaacaa aatgtaaaga tctgctgtgg tgcaatgctt gctgcgcgtc
     tgctgatgaa aagttcttga cccaaatgca gaaaatcaat agaatctgtg aaatcttcta
     taatcttaaa attagattaa agttctattt ttttgcccga gtttgtaacc acgggcgata
     aaaagtagct ttacgcctcg cacaccaata cacgaacaga aaaattatgc cggctgtaat
     atgagctcgg cgcgaaattt ctagatgacc ggttcttaga acatcaacct tgcatgtcca
     acaaatgctg gttaattaaa gacgtgcctt aacttaattt tcttggcaca cgtgcttatt
     tgaattcagt cttttgcact tgccatgcac acagccacac atatgtgaat ttgcgaattt
     gccactcatg catacactca tgtatgttcc atcatcgaga aaattcgaaa atcgtgaatc
     aaacttcggc atgaatcaaa tttcaaagag gtctttgttt ccacctggtt ctagaagttt
     cctttcgcgt gcttggatac ctatcttatg cataaacggt ttctgcacat gtaacttgaa
     cacatacaca cttgcaaaca tatgtatgta catatgcata ccctgaccac aaaattttca
     gcaaacttta gccgtacatc aaaccaccaa agagctgtgc tgttgtcaag gagaattttc
     ttccagaaag cttcaattag attgtttatc tgggggtgat gtacgcattg gacaacccta
     tgcgctctag aaacttccag taaatgttaa ctggatgtac aatgggtaca tccctaagcg
     tgcgagtgta tgcgtgttcg ctaactgtaa tgtatgtgtg ttcgtgtgcg aaagagaaaa
     ggatgagaag tctgccattt tgaaataaaa agattttgtg ctaggggggt ggggaaatat
     gattatcgaa aatgggcagt gaacaatgca gctgcatatt taatgagttg tgactaattc
     tcgtgtggta ttttcttgct cttccagatg cccgaagaag ctgagacttt cgcattccag
     gctgagattg ctcagcttat gtcgttgatc cggccaagct tggatggatt gcacgcaggt
     tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     agcgcgggga tctcatgctg gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt
     ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg
     tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt
     ggggaggcac gatggccgct ttggtcgatc gatgataaac ataaaaccaa ataaacaaca
     agcaaatgtg ttttaaaaat ctaacttctg agcgagtatt tattgggggg aataaacaat
     ctatgaatcg gattctttgc gcagcagctg ctcaatggcc tccaccgtgg acactccgtt
     ggttatcatt attatcttgt ttcgcgatcg agatcccttg tccaaagaaa cgtcgctctt
     tcgaagacct agaactttcg acagaaactt gaccagttcg gcgttagctt ctccctcgct
     gggcggagcg gcgatttgga cgcccactcc ttcaaagcca attcctgtga ttccgttctg
     cttagccccc ccggaattgg gtacccccac cgcggtggcg gccgctctag aactagtgga
     tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     cccggta
//