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ID   PSV2NEO    preliminary; circular DNA; SYN; 5729 BP.
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AC   IG0101; ATCC37149;
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DT   02-NOV-1992 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Vertebrate/E.coli plasmid vector pSV2neo - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RC   pSV2neo from pBR322 & SV40 & Tn5
RA   Garbern J.Y.;
RT   ;
RL   Unpublished (1992).
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RN   [2]
RC   pSV2neo from pBR322 & SV40 & Tn5
RA   Maekelae T.P.;
RT   ;
RL   Unpublished (1992).
RL   Maekelae T.P., Cancer Biology Lab., Dept. of Virology and Pathology,
RL   University of Helsinki, Haartmaninkatu 3, Helsinki 00290, FINLAND.
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RN   [3]
RC   pBR-neo from pRZ112 & pBR322
RC   pSV2-neo from pBR-neo & pSV2-betaG
RC   pSV3-neo from pSV2-neo
RC   pSV5-neo from pSV2-neo
RC   [SVGT10-betaG from SV40 & rabbit beta-globin gene]
RC   [SVGT10*-betaG from SVGT10-betaG]
RC   [pSV2 from pBR322 & SV40 early]
RC   [pSV2-betaG from pSV2 & rabbit beta-globin gene]
RC   [pSV2-neo-SVgpt from pSV2-neo & pSV2-gpt]
RA   Southern P.J., Berg P.;
RT   "Transformation of mammalian cells to antibiotic resistance with a
RT   bacterial gene under control of the SV40 early region promoter";
RL   J. Mol. Appl. Genet. 1:327-341(1982).
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RN   [4]
RC   pRZ326 from ColE1 & pBR322
RC   pRZ341 from ColE1 & Tn5-341
RA   Johnson R.C., Reznifkoff W.S.;
RT   "DNA sequences at the ends of transposon Tn5 required for
RT   transposition";
RL   Nature 304:280-282(1983).
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CC   ATCC size is 5600 bp.
CC   Provides dominant selectable marker for resistance to antibiotic G418
CC   in mammalian cell lines. (ATCC staff)
CC   Restriction digests of the clone give the following sizes (kb):
CC   PvuII--4.2, 0.74, 0.61, 0.36;  BamHI--5.8;  EcoRI--5.8;  PstI--2.45,
CC   1.40, 0.94 (doublet). (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pSV2neo)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (CLONTECH)(ATCC)
CC   HO (E.coli HB101)(E.coli)(human fibroblast or lymphoblast cells)
CC   HO (vertebrate cells)
CC   CP ()
CC   FN (helper)
CC   SE ()
CC   PA (pBR322)(SV40)(Tn5)
CC   BR ()
CC   OF (pSVneoB)
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322 4361bp
FT                   transposition
FT                   2. Tn5 5818bp
FT                   -> pRZ102 10179bp
FT                   1. pRZ102 remove pBR322 sequences
FT                   -> pRZ112
FT                   1. pRZ112 HincII-HincII 2499bp 188..2687, neo gene
FT                   BamHI linker 10bp nnggatccnn:BamHI linker 10bp
FT                   \ nnggatccnn
FT                   HindIII-BamHI 1493bp 1196..2689, neo/kan gene
FT                   2. pBR322 remove HindIII-BamHI 346bp 30..376, 4015bp
FT                   -> pBR-neo 5508bp
FT                   1. SV40 StuI-PvuII 323bp 5194..5243..273,
FT                   \ early promoter/HindIII-PvuII 344bp 5172..5243..273
FT                   2. pBR322 3045bp 324..3369, ori/amp gene
FT                   3. SV40 BamHI-MboI 237bp 2534..2771,
FT                   \ small t antigen [3370..3606]
FT                   4. SV40 MboI-? 521bp 2771..3292, polyA [3606..4127]
FT                   -> pSV2 4125bp
FT                   1. pSV2
FT                   2. rabbit, beta-globin gene
FT                   -> pSV2-betaG
FT                   1. pBR-neo HindIII-BamHI 1493bp 1196..2689, neo/kan
FT                   2. pSV2-betaG remove HindIII-BglII, beta-globin gene
FT                   -> pSV2-neo 5729bp"
FT   misc_binding    0..0
FT                   /note="SIT BamHI"
FT   rep_origin      complement(1..989)
FT                   /note="ORI SV40, from EcoRI-BclI fragment of SV40
FT                   (1782-2770), BclI site not retained."
FT   polyA_signal    827..827
FT                   /note="PLA SV40 early gene, addition site is at 3101,
FT                   from BglII-EcoRI fragment of psv2-gpt"
FT   misc_feature    990..1600
FT                   /note="SPL MboI-MboI SV40 intervening sequence
FT                   with splice acceptor (SV40 complement(4100..4710)),
FT                   from BglII-EcoRI fragment of psv2-gpt"
FT   misc_binding    0..0
FT                   /note="SIT HindIII"
FT   CDS             complement(1601..3093)
FT                   /note="ANT E. coli neomycin phosphotransferase II gene
FT                   (NPTII), neomycin resistance gene (neo),
FT                   from HindIII-BamHI fragment of transposon Tn5"
FT   misc_feature    complement(1950..1950)
FT                   /note="TER stop codon"
FT   misc_feature    complement(2742..2742)
FT                   /note="ATG"
FT   promoter        3094..3435
FT                   /note="PRO SV40 early gene (5171..5270 of SV40),
FT                   from PvuII-HindIII fragment of SV40"
FT   misc_binding    0..0
FT                   /note="SIT PvuII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322),
FT                   from EcoRI-PvuII fragment of pBR322"
FT   misc_feature    3434..5729
FT                   /note="PvuII-EcoRI of pBR322 (2066-4361 in pBR)"
FT   CDS             complement(>3434..<5729)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT PstI"
XX
SQ   Sequence 5729 BP; 1464 A; 1419 C; 1415 G; 1431 T; 0 other;
     aattcctttg cctaatttaa atgaggactt aacctgtgga aatattttga tgtgggaagc
     tgttactgtt aaaactgagg ttattggggt aactgctatg ttaaacttgc attcagggac
     acaaaaaact catgaaaatg gtgctggaaa acccattcaa gggtcaaatt ttcatttttt
     tgctgttggt ggggaacctt tggagctgca gggtgtgtta gcaaactaca ggaccaaata
     tcctgctcaa actgtaaccc caaaaaatgc tacagttgac agtcagcaga tgaacactga
     ccacaaggct gttttggata aggataatgc ttatccagtg gagtgctggg ttcctgatcc
     aagtaaaaat gaaaacacta gatattttgg aacctacaca ggtggggaaa atgtgcctcc
     tgttttgcac attactaaca cagcaaccac agtgcttctt gatgagcagg gtgttgggcc
     cttgtgcaaa gctgacagct tgtatgtttc tgctgttgac atttgtgggc tgtttaccaa
     cacttctgga acacagcagt ggaagggact tcccagatat tttaaaatta cccttagaaa
     gcggtctgtg aaaaacccct acccaatttc ctttttgtta agtgacctaa ttaacaggag
     gacacagagg gtggatgggc agcctatgat tggaatgtcc tctcaagtag aggaggttag
     ggtttatgag gacacagagg agcttcctgg ggatccagac atgataagat acattgatga
     gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     cctctacaaa tgtggtatgg ctgattatga tctctagtca aggcactata catcaaatat
     tccttattaa cccctttaca aattaaaaag ctaaaggtac acaatttttg agcatagtta
     ttaatagcag acactctatg cctgtgtgga gtaagaaaaa acagtatgtt atgattataa
     ctgttatgcc tacttataaa ggttacagaa tatttttcca taattttctt gtatagcagt
     gcagcttttt cctttgtggt gtaaatagca aagcaagcaa gagttctatt actaaacaca
     gcatgactca aaaaacttag caattctgaa ggaaagtcct tggggtcttc tacctttctc
     ttcttttttg gaggagtaga atgttgagag tcagcagtag cctcatcatc actagatggc
     atttcttctg agcaaaacag gttttcctca ttaaaggcat tccaccactg ctcccattca
     tcagttccat aggttggaat ctaaaataca caaacaatta gaatcagtag tttaacacat
     tatacactta aaaattttat atttacctta gagctttaaa tctctgtagg tagtttgtcc
     aattatgtca caccacagaa gtaaggttcc ttcacaaaga tccgggacca aagcggccat
     cgtgcctccc cactcctgca gttcgggggc atggatgcgc ggatagccgc tgctggtttc
     ctggatgccg acggatttgc actgccggta gaactccgcg aggtcgtcca gcctcaggca
     gcagctgaac caactcgcga ggggatcgag cccggggtgg gcgaagaact ccagcatgag
     atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag
     cccttgcgcc ctgagtgctt gcggcagcgt gaagcttttt gcaaaagcct aggcctccaa
     aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     gacacacatt ccacagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     gcgtatcacg aggccctttc gtcttcaag
//