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ID   PGEM5ZFM   preliminary; circular DNA; SYN; 3003 BP.
XX
AC   X65309; IG0144;
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DT   30-JUN-1988 (Rel. 3, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli phagemid vector pGEM-5Zf- - 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   pGEM-5Zf-, revised by [2]
RA   ;
RT   ;
RL   Submitted (23-MAR-1992) on tape to the EMBL Data Library by:
RL   Technical Services, Promega Corporation, 2800 Woods Hollow Road,
RL   Madison, WI 53711-5399, USA.
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RN   [2]
RP   1-3003
RC   pGEM-5Zf-
RA   Klock C.;
RT   ;
RL   Submitted (28-MAY-1993) on tape to the EMBL Data Library by:
RL   Technical Services, Promega Corporation, 2800 Woods Hollow Road,
RL   Madison, WI 53711-5399, USA.
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RN   [3]
RC   pGEM-5Zf- from pGEM series
RA   Lewis K.;
RT   ;
RL   Unpublished (1987).
RL   Lewis K., Promega.
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CC   The sequence is the strand that
CC   is secreted by cells harboring pGEM-3Zf(+) plasmids.
CC   NM (pGEM-5Zf-)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST (coding)
CC   TY (plasmid)
CC   SP (Promega)
CC   HO (E.coli JM109)
CC   CP ()
CC   FN (cloning)(transcription)(sequencing)(mutagenesis)
CC   SE (color blue/white)
CC   PA (pUC)(pGEM)
CC   BR (pGEM-5Zf+)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pGEM-3Z 2743bp
FT                   2. pUC18 250bp, f1 ori
FT                   -> pGEM-5Zf- 3003bp"
FT   misc_feature    1..1
FT                   /note="bacteriophage T7 transcription initiation
FT                   site"
FT   misc_binding    10..113
FT                   /note="MCS ApaI-AatII-SphI-NcoI-SacII-EcoRV-SpeI-
FT                   NotI-PstI-SalI-NdeI-SacI-BstXI-NsiI"
FT   misc_feature    126..126
FT                   /note="bacteriophage Sp6 transcription initiation
FT                   site"
FT   promoter        complement(121..143)
FT                   /note="PRO bacteriophage Sp6"
FT   misc_feature    complement(151..380)
FT                   /note="GEN E. coli lac operon"
FT   promoter        complement(185..201)
FT                   /note="PRO E. coli lac operator"
FT   primer_bind     161..177
FT                   /note="PRI bacteriophage M13 reverse"
FT   misc_feature    165..165
FT                   /note="E. coli lacZ start codon"
FT   rep_origin      complement(0..0)
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             complement(1322..2182)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    1875..1875
FT                   /note="SIT unique ScaI"
FT   misc_binding    1994..1994
FT                   /note="SIT unique XmnI"
FT   rep_origin      2368..2823
FT                   /note="ORI bacteriophage f1 intergenic region"
FT   misc_binding    2495..2495
FT                   /note="SIT unique NaeI"
FT   misc_feature    complement(2824..2984)
FT                   /note="GEN E. coli lac operon"
FT   primer_bind     2944..2960
FT                   /note="PRI bacteriophage M13 forward"
FT   promoter        join(2987..3003,1..6)
FT                   /note="PRO bacteriophage T7"
XX
SQ   Sequence 3003 BP; 726 A; 761 C; 746 G; 770 T; 0 other;
     gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggccgcggga tatcactagt
     gcggccgcct gcaggtcgac catatgggag agctcccaac gcgttggatg catagcttga
     gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     aaaaggccgc gttgctggcg tttttcgata ggctccgccc ccctgacgag catcacaaaa
     atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     acgttgttgg cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     ctgtgactgg tgagtactca accaagtcat tctgagaata ccgcgcccgg cgaccgagtt
     gctcttgccc ggcgtcaata cgggataata gtgtatgaca tagcagaact ttaaaagtgc
     tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     ttccgcgcac atttccccga aaagtgccac ctgtatgcgg tgtgaaatac cgcacagatg
     cgtaaggaga aaataccgca tcaggcgacg cgccctgtag cggcgcatta agcgcggcgg
     gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     gggggctccc tttagggttc cgatttagag ctttacggca cctcgaccgc aaaaaacttg
     atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     aaaatgagct gatttaacaa atatttaacg cgaattttaa caaaatatta acgtttacaa
     tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact
     ata
//