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ID   PEVVRF     preliminary; circular DNA; SYN; 4626 BP.
XX
AC   IG8023;
XX
DT   01-DEC-1994 (Rel. 10, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pEV-vrf - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RC   pRC1 from pBR322 & linker
RC   pRC2 from pBR322 & linker
RC   pRC23 from pRC2 & oligo & lambda, pL
RC   pEV-vrf1, pEV-vrf2, pEV-vrf3 from pRC23 & oligo
RA   Crowl R., Seamans C., Lomedico P., McAndrew S.;
RT   "Versatile expression vectors for high-level synthesis of cloned
RT   gene products in Escherichia coli";
RL   Gene 38:31-38(1985).
XX
CC   Created by Moore, July 1995, under contract with NCBI.
CC   NM (pEV-vrf)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pBR322)(lambda)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322 EcoRI 4361bp 4360..4360
FT                   S1 nuclease
FT                   BglII linker 8bp cagatctg
FT                   BglII-PstI 757bp 3612..4369
FT                   2. pBR322 EcoRI 4361bp 4360..4360
FT                   Klenow
FT                   BglII linker 8bp cagatctg
FT                   BglII-PstI 3613bp 4360..4361..3612
FT                   -> pRC2 4370bp [unique BglII next to EcoRI; 4350bp]
FT                   1. oligo BglII-HaeII 26bp, model RBS
FT                   \ agatctnnnnnnnnnnnnnnagcgct
FT                   2. lambda HpaI-BglII 450bp 35262..35712, pL
FT                   HaeIII-BglII 244bp 35468..35712, pL
FT                   -> BglII-BglII 270bp, pL/RBS
FT                   1. BglII-BglII 270bp, pL/RBS
FT                   2. pRC2 BglII 4364bp, linker
FT                   -> pRC23 4640bp [unique EcoRI in SD]
FT                   1. pRC23 remove small EcoRI-HindIII 31bp 4360..30
FT                   2. oligo EcoRI-HindIII 18bp aattcggatccatcgata
FT                   -> pEV-vrf1 4630bp [unique EcoRI]"
FT   -               1..4330
FT                   /note="pBR322 30..4359 4330bp
FT                   EcoRI = G^AATTC
FT                   \         ca"
FT   -               4331..4332
FT                   /note="ca 2bp
FT                   \ ca
FT                   \   gatct..."
FT   -               4333..4358
FT                   /note="gatctnnnnnnnnnnnnnnnagcgct 26bp
FT                   \ ...agcgct
FT                   HaeIII = GG^CC"
FT   -               4359..4602
FT                   /note="lambda 35468..35711 244bp
FT                   BglII = A^GATCT"
FT   -               4603..4608
FT                   /note="gatctg 6bp
FT                   \  gatctg
FT                   EcoRI = G^AATTC
FT                   \         aattc..."
FT   -               4609..4626
FT                   /note="aattcggatccatcgata 18bp
FT                   \  ...cgata
FT                   HindIII = A^AGCTT"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
XX
SQ   Sequence 4626 BP; 1035 A; 1266 C; 1193 G; 1117 T; 15 other;
     agctttaatg cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga
     aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag
     gcttggttat gccggtactg ccgggcctct tgcgggatat cgtccattcc gacagcatcg
     ccagtcacta tggcgtgctg ctagcgctat atgcgttgat gcaatttcta tgcgcacccg
     ttctcggagc actgtccgac cgctttggcc gccgcccagt cctgctcgct tcgctacttg
     gagccactat cgactacgcg atcatggcga ccacacccgt cctgtggatc ctctacgccg
     gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
     tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
     cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
     aggagtcgca taagggagag cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
     tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg
     ctcttaccag cctaacttcg atcactggac cgctgatcgt cacggcgatt tatgccgcct
     cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc
     tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg
     gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac
     tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag
     ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct
     cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     cgtcttcaag cagatctnnn nnnnnnnnnn nnagcgctcc aatgcttcgt ttcgtatcac
     acaccccaaa gccttctgct ttgaatgctg cccttcttca gggcttaatt tttaagagcg
     tcaccttcat ggtggtcagt gcgtcctgct gatgtgctca gtatcaccgc cagtggtatt
     tatgtcaaca ccgccagaga taatttatca ccgcagatgg ttatctgtat gttttttata
     tgaatttatt ttttgcaggg gggcattgtt tggtaggtga gagatctgaa ttcggatcca
     tcgata
//