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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.
XX
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).
XX
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
//