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ID   PMAMPF3    preliminary; circular DNA; SYN; 4354 BP.
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AC   M33637; ATCC77132;
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DT   27-JUL-1990 (Rel. 5, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pMAMPF3 - 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]
RP   1-4354
RC   pSZ1 from pMa5-14 & pIN-III-ompA3, NaeI fragment
RC   pMAMPF1 from pSZ1 & pMa5-14
RC   pMAMPF3 from pMAMPF1
RA   Szardenings M., Collins J.;
RT   "A plasmid optimised for protein design projects: pMAMPF";
RL   Gene 94:1-7(1990).
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RN   [2]
RC   pMAMPF3
RA   Collins J.;
RT   ;
RL   Submitted (09-APR-1990) to GenBank on computer-readable media by:
RL   Collins J., .
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RN   [3]
RC   pLK54 from phage fd terminator
RC   pcIwt from pKB280 & pcI857
RC   pMac5-8 from pLK54 & pBR325 & pEMBL8+ & pPLc28
RC   pMa5-8, pMc5-8 from pMac5-8
RC   pMa5-10, pMc5-10 from pcIwt & pBR325 & pEMBL8+ & pPLcAT14-betagal
RC   pMa5-14-betagal from pMa5-8 & pPLcAT14-betagal
RC   pMc5-14-betagal from pMc5-8 & pPLcAT14-betagal
RC   pMa5-14 from pMa5-14-betagal
RC   pMc5-14 from pMc5-14-betagal
RC   from pMa5-10
RC   from pMc5-10
RA   Stanssens P., Opsomer C., McKeown Y.M., Kramer W., Zabeau M.,
RA   Fritz H.J.;
RT   "Efficient oligonucleotide directed construction of mutations in
RT   expression vectors by the gapped duplex DNA method using alternating
RT   selectable markers";
RL   Nucleic Acids Res. 17:4441-4454(1989).
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CC   Expression vector permitting production of a fusion protein and
CC   containing signal sequences for protein export.  Also permits
CC   production of ssDNA and contains priming sites useful for sequencing.
CC   Encodes the ompA leader.  Contains a chloramphenical acetyltransferase
CC   gene with an amber mutation useful for gapped-duplex mutagenesis
CC   experiments.
CC   Expression can be induced by a temperature shift in hosts providing a
CC   temperature-sensitive lambda repressor, or by batch transduction in a
CC   growing culture to repressor-negative hosts.
CC   The order of the major features in this phagemid is:lambdaPL promoter
CC   - ompA leader - NaeI site - MCS - priming site - fd ori - pMB1 ori -
CC   bla - cat (am).
CC   Restriction digests of the clone give the following sizes (kb):
CC   SalI--4.3; PstI--2.3, 2.0; XbaI--4.3; EcoRI--4.3; HindIII--4.3. (ATCC
CC   staff)
CC   A (GT)n repeat. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pMAMPF3)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)
CC   CP ()
CC   FN (protein design)
CC   SE ()
CC   PA (pBR325)(pMAC5-8)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322
FT                   2. phage fd Sau3A/MboI-Sau3A/MboI 332bp 1382..1714,
FT                   \ terminator [338bp]
FT                   \ MboI= 1382 1714 2221 5646
FT                   -> pBRT7 4400bp
FT                   1. pBR322 remove EcoRI-PvuII 2068bp
FT                   \ 4360..4361..2067, 2293bp
FT                   2. pBRT7 EcoRI*-EcoRI* 99bp, fd terminator
FT                   -> pLK12 2392bp
FT                   1. pPLc28 2720bp
FT                   2. pLK12 2392bp
FT                   -> pLK54 5100bp
FT                   1. pLK54 100bp, MCS/phage fd term 28bp in tandem
FT                   2. pEMBL8+ 456bp, f1 ori
FT                   3. pPLc28 2720bp, ori/amp gene
FT                   4. pBR325 EcoRI 5994bp 4778..4778, cat gene
FT                   mutagenesis
FT                   BalI 5994bp 5047..5047, cat gene
FT                   mutagenesis
FT                   NcoI 5994bp 5079..5079, cat gene
FT                   mutagenesis
FT                   -> pMac5-8 3803bp
FT                   1. pMac5-8 3803bp
FT                   amber mutation in cat gene at PvuII
FT                   -> pMa5-8 3803bp [no PvuII]
FT                   1. pPLc28 2720bp
FT                   2. lambda, pL promoter
FT                   -> pPLcAT14-betagal 3000bp
FT                   1. pMa5-8 remove BssHII-BamHI, MCS/3800bp
FT                   2. pPLcAT14-betagal BssHII-BamHI, pL/lacZ gene/3000bp
FT                   -> pMa5-14-betagal 6800bp
FT                   1. pMa5-14-betagal remove BamHI-BamHI, lacZ/4145bp
FT                   -> pMa5-14 4145bp
FT                   1. pIN-III-A3 7400bp
FT                   2. E. coli, ompA3 gene
FT                   -> pIN-III-ompA3
FT                   1. pIN-III-ompA3
FT                   -> pIN-III-ompA3-NaeI [NaeI at end of leader]
FT                   1. pMa5-14 XbaI-PstI 4000bp
FT                   calf intestinal alkaline phosphatase
FT                   2. pIN-III-ompA3-NaeI large XbaI-PstI
FT                   -> pSZ1
FT                   1. pSZ1 small BamHI-PstI 100bp
FT                   2. pMa5-14 BamHI-PstI 2100bp
FT                   3. pMa5-14 BamHI-PstI 2200bp
FT                   -> pMAMPF1 4354bp
FT                   1. pMAMPF1 NaeI 4354bp 4184..4184
FT                   mutagenesis
FT                   -> pMAMPF3 4354bp"
FT   sig_peptide     4121..>4183
FT                   /note="E. coli OmpA signal sequence"
FT   misc_binding    4181..4186
FT                   /note="SIT unique NaeI"
FT   misc_feature    4184..4308
FT                   /note="stuffer fragment, from pBR325 5900..5996..29"
FT   misc_binding    0..0
FT                   /note="MCS HindIII-SalI-PstI-HindIII-XbaI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage fd"
FT   promoter        0..0
FT                   /note="PRO bacteriophage lambda pL"
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SQ   Sequence 4354 BP; 1152 A; 1054 C; 1012 G; 1136 T; 0 other;
     aattcacctc gaaagcaagc tgataaaccg atacaattaa aggctccttt tggagccttt
     ttttttggag attttcaacg tgaaaaaatt attattcgca attccaagct aattcacctc
     gaaagcaagc tgataaaccg atacaattaa aggctccttt tggagccttt ttttttggag
     attttcaacg tgaaaaaatt attattcgca attccaagct ctgcctcgcg cgtttcggtg
     atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     cggatgcaga tcacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     cctttctcgc cacgttcgcc agctttcccc gtcaagctct aaatcggggg ctccctttag
     ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttga tctgcgctcg
     gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcagacag
     ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgactcccc gcgcgcgatg
     ggtcgaattt gctttcgaaa aaaaagcccg ctcattaggc gggctaaaaa aaagcccgct
     cattaggcgg gctcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacagacgg
     catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     ccatagtgaa aacgggggcg aagaagttgt ccatattcgc cacgtttaaa tcaaaactgg
     tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc
     cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
     tttattcgaa gacgaaaggg catcgcgcgc ggggaattgg ccacgatgcg tccggcgtag
     aggatctctc acctaccaaa caatgccccc ctgcaaaaaa taaattcata taaaaaacat
     atagataacc atctgcggtg ataaattatc tctggcggtg ttgacataaa taccactggc
     ggtgatactg agcacatcag caggacgcac tgaccaccat gaaggtgacg ctcttaaaat
     taagccctga agaagggcag cattcaaagc agaaggcttt ggggtgtgtg atacgaaacg
     aagcattgga attctacaac ttgcttggat tcctacaaag aagcagcaat tttcagtgtc
     agaagtcgac caaggaggtc tagataacga ggcgcaaaaa atgaaaaaga cagctatcgc
     gattgcagtg gcactggctg gtttcgctac cgtagcgcag gccggcgata atggcctgct
     tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     ttccgaatac cgcaagcgac aggccatgtt tgacagctta tcatcgataa gcttggatcc
     gtcgacctgc agccaagctt ggtctagagg tcga
//