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ID   PMALC      preliminary; circular DNA; SYN; 6148 BP.
XX
AC   IG1997; 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-c [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
RN   [2]
RC   pCG806fX from pCG806 & oligo, factor Xa
RA   Maina C.V., Riggs P.D., Grandea A.G., Slatko B.E., Moran L.S.,
RA   Tagliamonte J.A., McReynolds L.A., Guan C.D.;
RT   "An Escherichia coli vector to express and purify foreign proteins
RT   by fusion to and separation from maltose-binding protein";
RL   Gene 74:365-373(1988).
XX
RN   [3]
RC   [from none]
RA   Riggs P.D.;
RT   ;
RL   Current Protocols in Molecular Biology 1992:0-0(1992).
RL   Ausubel F.M., ed., Wiley, New York.
XX
CC   old NEB vector.
CC   NM (pMAL-c [tm])
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (expression)
CC   SE ()
CC   PA (pKK233-2)
CC   BR (pMAL-c2 [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-c 6148bp"
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    2729..2729
FT                   /note="start E. coli lacZ-alpha sequence (CA CTG GCC)"
FT   misc_feature    2895..2895
FT                   /note="start E. coli rrnB terminator DNA fragment"
FT   misc_feature    2908..2908
FT                   /note="TER first in-frame stop codon for E. coli
FT                   lacZ-alpha"
FT   misc_feature    3321..3321
FT                   /note="start pBR322 sequence"
FT   CDS             3412..3412
FT                   /note="E. coli beta-lactamase start codon (ATG)"
FT   misc_feature    4270..4270
FT                   /note="E. coli beta-lactamase stop codon (TAA)"
FT   misc_feature    6145..6145
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 6148 BP; 1522 A; 1563 C; 1645 G; 1418 T; 0 other;
     ccgacaccat cgaatggcgc 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 agctggcgaa 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
     gtacccggcc ggggatccat cgagggtagg cctgaattca gtaacctaac cctcgatgga
     tcctctagag tcgacctgca ggcaagcttg gcactggccg tcgttttaca acgtcgtgac
     tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     ggcgaatggc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     ttttgcgttt ctacaaactc tttttgttta tttttctaaa tacattcaaa tatgtatccg
     ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     cctgtttggt cacttgatgc ctccgtgtaa gggggaattt ctgttcatgg gggtaatgat
     accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     tggccaggac ccaacgctgc ccgaaatt
//