pCAP01

LOCUS       pCAP01                9022 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  The function of this material is to facilitate the capture and 
            expression of bacterial gene clusters..
ACCESSION   .
VERSION     .
KEYWORDS    pCAP01
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9022)
  AUTHORS   Yamanaka K, Reynolds KA, Kersten RD, Ryan KS, Gonzalez DJ, Nizet V, 
            Dorrestein PC, Moore BS
  TITLE     Direct cloning and refactoring of a silent lipopeptide biosynthetic 
            gene cluster yields the antibiotic taromycin A.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Feb 4;111(5):1957-62. doi: 
            10.1073/pnas.1319584111. Epub 2014 Jan 21.
  PUBMED    24449899
REFERENCE   2  (bases 1 to 9022)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9022
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    84..587
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     629..647
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(685..707)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     807..826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        847..1126
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1127..1801
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(2041..2062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2309..2897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2798..2817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3062..3079
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3293..3427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3317..3336)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3371..3390
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3852..3872)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4003..4068
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4488..5282)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4599..4618)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5209..5228
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(6066..6437)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6470..6579
                     /note="incP origin of transfer"
     CDS             complement(6817..6828)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    6899..6998
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             7008..8825
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RAPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
ORIGIN
        1 acttgaggta cctgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
       61 aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt
      121 taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt
      181 tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc
      241 ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac
      301 cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt
      361 taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt
      421 taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt
      481 tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt
      541 ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac
      601 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
      661 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      721 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      781 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      841 tagatcacga cattactata tatataatat aggaagcatt taatagacag catcgtaata
      901 tatgtgtact ttgcagttat gacgccagat ggcagtagtg gaagatattc tttattgaaa
      961 aatagcttgt caccttacgt acaatcttga tccggagctt ttcttttttt gccgattaag
     1021 aattaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt ggtgactatt
     1081 gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt ctgttattaa
     1141 tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca cagaggccgc
     1201 agaatgtgct ctagattccg atgctgactt gctgggtatt atatgtgtgc ccaatagaaa
     1261 gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag catataaaaa
     1321 tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta aggaggatgt
     1381 tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag atgagtcgtg
     1441 gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat ttccaaaaga
     1501 ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc ccttgtttga
     1561 ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg actgggttgg
     1621 aaggcaagag agccccgaaa gcttacattt tatgttagct ggtggactga cgccagaaaa
     1681 tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg gaggtgtgga
     1741 gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg ctaagaaata
     1801 ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgccgat ctatgcggtg
     1861 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     1921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2101 accgtctatc agggcgatgg cccactacgt gaacctaact gtcagaccaa gtttactcat
     2161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cctaccgggt
     2581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2941 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3001 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3061 cagcgagtca gtgagcgagg aagcggaaga gcgctgactt ccgcgtttcc agactttacg
     3121 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     3181 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     3241 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt cagatccaga
     3301 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3361 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3421 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3481 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atctctagtc
     3541 aaggcactat acatcaaata ttccttatta acccctttac aaattaaaaa gctaaaggta
     3601 cacaattttt gagcatagtt attaatagca gacactctat gcctgtgtgg agtaagaaaa
     3661 aacagtatgt tatgattata actgttatgc ctacttataa aggttacaga atatttttcc
     3721 ataattttct tgtatagcag tgcagctttt tcctttgtgg tgtaaatagc aaagcaagca
     3781 agagttctat tactaaacac agcatgactc aaaaaactta gcaattctga aggaaagtcc
     3841 ttggggtctt ctacctttct cttctttttt ggaggagtag aatgttgaga gtcagcagta
     3901 gcctcatcat cactagatgg catttcttct gagcaaaaca ggttttcctc attaaaggca
     3961 ttccaccact gctcccattc atcagtttcc ataggttgga atctaaaata cacaaacaat
     4021 tagaatcagt agtttaacac attatacact taaaaatttt atatttacct tagagcttta
     4081 aatctctgta ggtagtttgt ccaattatgt cacaccacag aagtaaggtt ccttcacaaa
     4141 gatccggacc aaagcggcca tcgtgcctcc ccactcctgc agttcggggg catggatgcg
     4201 cggatagccg ctgctggttt cctggatgcc gacggatttg cactgccggt agaactccgc
     4261 gaggtcgtcc agcctcaggc agcagctgaa ccaactcgcg aggggatcga gcccggggtg
     4321 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     4381 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     4441 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     4501 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     4561 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     4621 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     4681 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     4741 ccgtcgggca tccgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     4801 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     4861 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     4921 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     4981 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     5041 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     5101 tggagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     5161 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     5221 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     5281 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
     5341 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     5401 gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc
     5461 catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag
     5521 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     5581 gtgttccgct tcctttagca gcccttgcgc cctgagtgct tgcggcagcg tgagagctca
     5641 tgagcggaga acgagatgac gttggagggg caaggtcgcg ctgattgctg gggcaacacg
     5701 tggagcggat cggggattgt ctttcttcag ctcgctgatg atatgctgac gctcaatgcc
     5761 gtttggcctc cgactaacga aaatcccgca tttggacggc tgatccgatt ggcacggcgg
     5821 acggcgaatg gcggagcaga cgctcgtccg ggggcaatga gatatgaaaa agcctgaact
     5881 caccgcgacg tatcgggccc tggccagcta gctagagtcg acctgcaggt ccccggggat
     5941 cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga
     6001 tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa
     6061 aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc
     6121 tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
     6181 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
     6241 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
     6301 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc
     6361 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
     6421 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
     6481 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
     6541 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
     6601 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
     6661 tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagatccgt
     6721 cgacctgcag gcatgcaagc tctagcgatt ccagacgtcc cgaaggcgtg gcgcggcttc
     6781 cccgtgccgg agcaatcgcc ctgggtgggt tacacgacgc ccctctatgg cccgtactga
     6841 cggacacacc gaagccccgg cggcaaccct cagcggatgc cccggggctt cacgttttcc
     6901 caggtcagaa gcggttttcg ggagtagtgc cccaactggg gtaacctttg agttctctca
     6961 gttgggggcg tagggtcgcc gacatgacac aaggggttgt gaccggggtg gacacgtacg
     7021 cgggtgctta cgaccgtcag tcgcgcgagc gcgaaaattc gagcgcagca agcccagcga
     7081 cacagcgtag cgccaacgaa gacaaggcgg ccgaccttca gcgcgaagtc gagcgcgacg
     7141 ggggccggtt caggttcgtc gggcatttca gcgaagcgcc gggcacgtcg gcgttcggga
     7201 cggcggagcg cccggagttc gaacgcatcc tgaacgaatg ccgcgccggg cggctcaaca
     7261 tgatcattgt ctatgacgtg tcgcgcttct cgcgcctgaa ggtcatggac gcgattccga
     7321 ttgtctcgga attgctcgcc ctgggcgtga cgattgtttc cactcaggaa ggcgtcttcc
     7381 ggcagggaaa cgtcatggac ctgattcacc tgattatgcg gctcgacgcg tcgcacaaag
     7441 aatcttcgct gaagtcggcg aagattctcg acacgaagaa ccttcagcgc gaattgggcg
     7501 ggtacgtcgg cgggaaggcg ccttacggct tcgagcttgt ttcggagacg aaggagatca
     7561 cgcgcaacgg ccgaatggtc aatgtcgtca tcaacaagct tgcgcactcg accactcccc
     7621 ttaccggacc cttcgagttc gagcccgacg taatccggtg gtggtggcgt gagatcaaga
     7681 cgcacaaaca ccttcccttc aagccgggca gtcaagccgc cattcacccg ggcagcatca
     7741 cggggctttg taagcgcatg gacgctgacg ccgtgccgac ccggggcgag acgattggga
     7801 agaagaccgc ttcaagcgcc tgggacccgg caaccgttat gcgaatcctt cgggccccgc
     7861 gtattgcggg cttcgccgct gaggtgatct acaagaagaa gccggacggc acgccgacca
     7921 cgaagattga gggttaccgc attcagcgcg acccgatcac gctccggccg gtcgagcttg
     7981 attgcggacc gatcatcgag cccgctgagt ggtatgagct tcaggcgtgg ttggacggca
     8041 gggggcgcgg caaggggctt tcccgggggc aagccattct gtccgccatg gacaagctgt
     8101 actgcgagtg tggcgccgtc atgacttcga agcgcgggga agaatcgatc aaggactctt
     8161 accgctgccg tcgccggaag gtggtcgacc cgtccgcacc tgggcagcac gaaggcacgt
     8221 gcaacgtcag catggcggca ctcgacaagt tcgttgcgga acgcatcttc aacaagatca
     8281 ggcacgccga aggcgacgaa gagacgttgg cgcttctgtg ggaagccgcc cgacgcttcg
     8341 gcaagctcac tgaggcgcct gagaagagcg gcgaacgggc gaaccttgtt gcggagcgcg
     8401 ccgacgccct gaacgccctt gaagagctgt acgaagaccg cgcggcaggc gcgtacgacg
     8461 gacccgttgg caggaagcac ttccggaagc aacaggcagc gctgacgctc cggcagcaag
     8521 gggcggaaga gcggcttgcc gaacttgaag ccgccgaagc cccgaagctt ccccttgacc
     8581 aatggttccc cgaagacgcc gacgctgacc cgaccggccc taagtcgtgg tgggggcgcg
     8641 cgtcagtaga cgacaagcgc gtgttcgtcg ggctcttcgt agacaagatc gttgtcacga
     8701 agtcgactac gggcaggggg cagggaacgc ccatcgagaa gcgcgcttcg atcacgtggg
     8761 cgaagccgcc gaccgacgac gacgaagacg acgcccagga cggcacggaa gacgtagcgg
     8821 cgtagcgaga cacccgggaa gcctgatcta cgtctgtcga gaagtttctg atcgaaaagt
     8881 tcgacagcgt ctccgacctg atgcagctct cgcagggcga agaatctcgt gctttcagct
     8941 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     9001 aagatcgact agtaacctcg ag
//