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ID   PMALC2     preliminary; circular DNA; SYN; 6646 BP.
XX
AC   IG1998; M24051; M24052; M21655; M21656;
XX
DT   01-JUN-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   E. coli plasmid vector pMAL-c2 [tm] - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RC   pPL-5A from malE gene
RC   pCG806 from pUC18 & pmalE, malE gene
RC   pCG150 from pUC18 & pPL-5A, malE gene
RC   pCG325 from pCG150 & pMLB1034
RC   pPL-6A' from pHI28 & pmalE & pCH2
RC   pCG228 from pUC18 & pGW4400
RC   pCG410 from pCG228 & pCG150
RC   pCG828 from pCG228 & pCG806
RC   pCG1001 from pACYC184 & modification methylase
RA   Guan C.D., Li P., Riggs P.D., Inouye H.;
RT   "Vectors that facilitate the expression and purification of
RT   foreign peptides in Escherichia coli by fusion to maltose-binding
RT   protein";
RL   Gene 67:21-30(1988).
XX
CC   NM (pMAL-c2 [tm])
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (NEB)
CC   HO (E.coli)
CC   CP ()
CC   FN (expression)
CC   SE ()
CC   PA (pKK233-2)
CC   BR (pMAL-cRI [tm])
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. E. coli, malE gene
FT                   -> pMAL-c2 6646bp"
FT   promoter        15..46
FT                   /note="PRO E. coli lacI gene"
FT   misc_feature    81..83
FT                   /note="ATG E. coli lacI gene start codon (GTG)"
FT   misc_feature    1161..1163
FT                   /note="TER E. coli lacI gene termination codon (TGA)"
FT   misc_feature    1205..1205
FT                   /note="begin Ptac DNA fragment"
FT   promoter        1406..1433
FT                   /note="PRO E. coli tac"
FT   misc_feature    1478..1478
FT                   /note="begin E. coli malE DNA fragment"
FT   misc_feature    1528..1528
FT                   /note="ATG E. coli malE start codon (ATG)"
FT   misc_feature    2587..2610
FT                   /note="PRI E. coli malE primer #1237"
FT   misc_feature    2629..2629
FT                   /note="MCS, also factor Xa site"
FT   misc_feature    2735..2735
FT                   /note="start E. coli lacZ-alpha sequence (CA CTG GCC)"
FT   misc_feature    2901..2901
FT                   /note="start E. coli rrnB terminator DNA fragment"
FT   misc_feature    2914..2914
FT                   /note="TER first in-frame stop codon for E. coli
FT                   lacZ-alpha"
FT   misc_feature    3327..3327
FT                   /note="start pBR322 sequence"
FT   CDS             3418..3418
FT                   /note="E. coli beta-lactamase start codon (ATG)"
FT   misc_feature    4276..4276
FT                   /note="E. coli beta-lactamase stop codon (TAA)"
FT   rep_origin      4320..4833
FT                   /note="ORI bacteriophage f1 origin"
FT   misc_feature    6646..6646
FT                   /note="end pBR322 sequence"
FT   misc_binding    7..7
FT                   /note="SIT unique PflMI"
FT   CDS             81..1161
FT                   /note="REP E. coli lacIq repressor gene"
FT   misc_binding    431..431
FT                   /note="SIT unique MluI"
FT   misc_binding    612..612
FT                   /note="SIT unique BstEII"
FT   misc_binding    638..638
FT                   /note="SIT unique ApaI"
FT   misc_binding    879..879
FT                   /note="SIT unique EcoRV"
FT   misc_binding    935..935
FT                   /note="SIT unique HpaI"
FT   misc_binding    1070..1070
FT                   /note="SIT unique NarI"
FT   promoter        1406..1433
FT                   /note="PRO E. coli tac (trp -35 and lacUV5 -10)"
FT   CDS             1528..2661
FT                   /note="GEN E. coli malE gene,
FT                   codes for maltose binding protein (MBP),
FT                   has no malE signal sequence"
FT   misc_binding    1893..1893
FT                   /note="SIT unique BsiWI"
FT   misc_binding    1962..1962
FT                   /note="SIT unique BglII"
FT   misc_binding    2217..2217
FT                   /note="SIT unique BsmI"
FT   misc_binding    2388..2388
FT                   /note="SIT unique Bpu1102I"
FT   misc_binding    2564..2564
FT                   /note="SIT unique NcoI"
FT   misc_binding    2704..2809
FT                   /note="MCS unique SacI-XmnI-EcoRI-BamHI-XbaI-SalI-
FT                   PstI-HindIII"
FT   misc_binding    2763..2764
FT                   /note="SIT factor Xa"
FT   CDS             2810..2968
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ)
FT                   alpha peptide"
FT   terminator      2989..2989
FT                   /note="TER E. coli beta-galactosidase gene (lacZ)
FT                   stop codon"
FT   terminator      0..0
FT                   /note="TER E. coli rrnB gene T1"
FT   terminator      0..0
FT                   /note="TER E. coli rrnB gene T2"
FT   CDS             3493..4351
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    3797..3797
FT                   /note="SIT unique ScaI"
FT   misc_binding    4259..4259
FT                   /note="SIT unique Pfl108I"
FT   misc_binding    4275..4275
FT                   /note="SIT unique Eam1105I"
FT   rep_origin      4395..4908
FT                   /note="ORI bacteriophage M13 ori-, from pZ150"
FT   misc_binding    4450..4450
FT                   /note="SIT unique SwaI"
FT   misc_binding    4672..4672
FT                   /note="SIT unique DraIII"
FT   misc_binding    4778..4778
FT                   /note="SIT unique NaeI"
FT   rep_origin      >4943..<5663
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      5607..5607
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322) start"
FT   misc_binding    5785..5785
FT                   /note="SIT unique SapI"
FT   misc_binding    5841..5841
FT                   /note="SIT unique NdeI"
FT   misc_binding    5892..5892
FT                   /note="SIT unique AccI"
FT   misc_binding    5916..5916
FT                   /note="SIT unique Tth111I"
FT   CDS             complement(>6000..<6378)
FT                   /note="REP E. coli ROP gene"
FT   misc_binding    6137..6137
FT                   /note="SIT unique BspGI"
FT   misc_binding    6557..6557
FT                   /note="SIT unique Bpu10I"
FT   misc_binding    6694..6694
FT                   /note="SIT MscI"
XX
SQ   Sequence 6646 BP; 1693 A; 1671 C; 1768 G; 1514 T; 0 other;
     ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     gcacttcacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
     attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg
     ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc
     ggatcctcta gagtcgacct gcaggcaagc ttggcactgg ccgtcgtttt acaacgtcgt
     gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     aatggcgaat ggcagcttgg ctgttttggc ggatgagata agattttcag cctgatacag
     attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     cctttttgcg tttctacaaa ctctttttgt ttatttttct aaatacattc aaatatgtat
     ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     gaacgttctc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa
     ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt
     gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg
     cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     tgttaagggc ggttttttcc tgtttggtca cttgatgcct ccgtgtaagg gggaatttct
     gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gaaatt
//