LOCUS       40924_49247        6061 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD053, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6061)
  AUTHORS   Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, 
            Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
  TITLE     Broad-host-range vector system for synthetic biology and 
            biotechnology in cyanobacteria
  JOURNAL   Nucleic Acids Res. (2014) In press
  PUBMED    25074377
REFERENCE   2  (bases 1 to 6061)
  AUTHORS   Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B, 
            Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2014) Division of Biological Sciences, University 
            of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE   3  (bases 1 to 6061)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6061)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2014) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JUN-2014) Division of Biological Sciences, University of 
            California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    405..410
                     /label=ZraI
                     /note="ZraI"
     misc_feature    408..3776
                     /note="CVD-CYA; device for the GC-adaptor assembly: 
                     cyanobacterial replicons"
     misc_feature    408..428
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    429..434
                     /label=XbaI
                     /note="XbaI"
     misc_feature    496..501
                     /label=XbaI
                     /note="XbaI"
     CDS             806..1357
                     /codon_start=1
                     /product="orfC"
                     /label=orfC
                     /note="orfC-pDU1; derived from pAM0505;
                     integrase/recombinase plasmid stability; similar to INSD 
                     accession AAM76170; Nostoc sp. PCC7524"
                     /protein_id="AII99682.1"
                     /translation="MKVNGNGRAKILTSDELRRLFSDGFTTPRDRVLFGICLFTGCRVS
                     EALALQTTDIKGETLTFRKSTTKGKLKTRVVDIQPGLAALMADYHPKPGTLFPGMRGVS
                     DRLTRYAADKILRDAAKRIGLEGISTHSFRRTALNQMSSAGIPLRHIQEISGHNDLGTL
                     QRYLEVTPEQRRKAVSVIGF"
     misc_feature    923..928
                     /label=AgeI
                     /note="AgeI"
     misc_feature    1264..1269
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    1456..1461
                     /label=XbaI
                     /note="XbaI"
     regulatory      complement(1872..1899)
                     /note="rho-ITTS; rho-ITTS; rho-independent transcriptional 
                     termination signal, downstream of repA in pDU1, from 
                     pAM505; Nostoc sp. PCC7524"
                     /regulatory_class="terminator"
     CDS             complement(1978..3099)
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDU1; derived from pAM0505; Nostoc sp. PCC7524"
                     /protein_id="AII99683.1"
                     /translation="MISPESLNTNVSLTLLTRELQTEIILGTYTVRVHTRIGREPCARL
                     WYLCRALDKDGSGHLTLPLPVVQTFLDCSDKSVYRWLQDGKKIGAFRRYKIKAGMITVY
                     LGGMFQVCYNLNLKRWGDVAVVPLVQVLSDLRSLTTGIVTQSFQQKSRYAANRQLKPEY
                     RKLFGAPHPNELVKDTRQSSLKSPEGEVPCVLHISSSRIFVSKSFIHYGTSQKAVSCEL
                     GIHKRTVRRHQKQLGMNRRQLCQAKIEYNQLRHARNNDASEFWAFTGTKTDIGYQVMGD
                     AVVFSDGIAPGAKKRQPNTYQIDATEFDGRLFKVGDKVFMNRCNIYREQFTLTTMSAAR
                     RKYHFKLSQCHFSENRAGRVGNRFVIGCHSGEI"
     misc_feature    2278..2283
                     /label=EcoRV
                     /note="EcoRV"
     regulatory      complement(3100..3749)
                     /note="PrepA; PrepA; upstream region of repA of pDU1
                     including promoter and ribosomal binding site,derived from 
                     pAM505; Nostoc sp. PCC7524"
                     /regulatory_class="promoter"
     misc_feature    3345..3350
                     /label=MfeI
                     /note="MfeI"
     misc_feature    3750..3755
                     /label=MfeI
                     /note="MfeI"
     misc_feature    3756..3776
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    3774..3779
                     /label=ZraI
                     /note="ZraI"
     misc_feature    3798..3803
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(3840..3856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3864..3880)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3888..3918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3933..3954)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4242..4830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5004..5861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5862..5966)
                     /label=AmpR promoter
     misc_feature    5992..5997
                     /label=ZraI
                     /note="ZraI"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgaggacgtc gggggccccc
      421 ggggggattc tagattcagg cagaaggaca agctaccggg caagacagaa agaatataga
      481 caatagtata gacaatctag acaaattgga ggcaacccgc gatcgcaccc tcaataagct
      541 aaaaatgggt aggcagtcag ccgccgggaa agccatcgac gcgtttatca aagagttgct
      601 ttcttcagga gacaacataa gctgaagtta tcaaaattct gtccttacgt cgaaagcctg
      661 attttaccgt gcaacgattg ataagcttgg ctaaactagc actggctttc aacagaaagc
      721 atacgaagaa tcaatagata tagccaccaa ttccacaaaa tgcagataac gtgtagagta
      781 ttggaatgct taatctgtaa gggttatgaa ggttaacggc aacggacgag ccaaaatact
      841 cacctccgac gaactcaggc gactgtttag cgacggattc accacaccgc gcgatcgcgt
      901 tttgtttggc atctgtctat tcaccggttg ccgcgttagt gaagctctag cactccaaac
      961 aacggacatt aaaggcgaaa cactaacctt taggaagtct accaccaaag ggaaactcaa
     1021 aacccgcgtg gttgacatcc agccaggact agccgcactc atggctgact atcaccccaa
     1081 accgggaacc ctgttccctg gcatgagggg agtcagcgat aggctcacgc gatacgcggc
     1141 ggataaaatc ttgcgcgatg cagccaaaag aatcgggcta gaaggcatca gtacccacag
     1201 tttccgccgt actgccctca accaaatgtc tagcgccggt atcccgttgc gacacattca
     1261 agagatatcc ggtcacaatg accttggcac actgcaacgc tatcttgaag ttacacccga
     1321 acagcgacgc aaagctgtat ccgtgattgg cttctaatgt acgccaacgc tgtttagacc
     1381 cctatgggtg ctaaaaaaag acgcagccta aacacacgct ctacacttga ggatactttt
     1441 aaagtatcca tcggttctag aactctgcac acgttccgga ctttggaaac gttatacctt
     1501 tccctgtgtt gcagaatgct gcaatatttc ttcgacaagt taacttgtga ctggtttaat
     1561 attttctcaa attgccccaa aacaacacgc ctaaatcctt agacgtttct gtggaaacct
     1621 attaggtttt tatcgccgtt gttttagtgg taaacccaaa gggtttgtat attcttgtat
     1681 gaagttcgac tctgagggtt aagaagaatg gctcgccgaa ttttttacaa gtggaaaccg
     1741 attaaaggtt aagggtcaat cgggacgatg aatattttct aattgtgacc ttctccatct
     1801 aataagcttt ctttggggtt aaggtcgaag aaagtactac gcatgatctg catacgatct
     1861 ctattgccaa aaagccgcga ccctataggc tctcggtcat gctgcactag ttcgtgtcga
     1921 tcactatact ggttgccgca gcatttcacg ctaaaaaaaa attcttaaaa atgtccttca
     1981 tatctcgcca gagtggcaac ctattacaaa acggttgcct acccgaccgg ctcgattttc
     2041 gctgaagtgg cactgtgaca gtttgaaatg gtacttccgc cgtgctgctg acatcgttgt
     2101 tagggtgaat tgttcgcggt agatgttgca ccgattcatg aacaccttgt cacccacttt
     2161 gaataatcga ccgtcaaatt cagtcgcgtc aatttggtaa gtgttgggct gtctcttttt
     2221 ggctccaggg gcaatgccat cagaaaacac aaccgcgtca cccataactt gataaccgat
     2281 atcagttttg gttccagtga aagcccaaaa ttcagacgcg tcattattcc gagcgtgccg
     2341 gagttgattg tactcaattt tggcttggca aagttgacgg cgattcatgc ccagctgctt
     2401 ttgatgtcgt cgcactgtgc gcttgtgaat acccaactca cagctgacag ctttttgaga
     2461 tgtaccatag tggatgaaac tttttgagac gaatatccgc gacgaactaa tgtgaagtac
     2521 acaaggtact tccccctctg gcgatttaag agaggattgc cttgtgtcct tcactagctc
     2581 gttcgggtgt ggcgctccaa aaagttttct gtactctggt ttaagttgtc tgttggccgc
     2641 atagcggctc ttttgttgaa agctttgtgt gactatgcca gtggtcagtg agcgtaaatc
     2701 gcttaacact tggactaaag gcactactgc aacatcaccc catcttttta aatttaggtt
     2761 gtaacaaact tgaaacatac cgcccaagta gacggttatc attcctgctt taattttgta
     2821 gcggcggaat gctcctattt tttttccatc ctgtaaccaa cggtaaacag acttatcact
     2881 acaatctaag aacgtctgta ctacaggcaa tggcaatgtt aaatgaccag acccatcctt
     2941 atcaagcgct cgacacaaat accacaaccg cgcacaaggt tctcgaccaa tgcgagtgtg
     3001 taccctgacc gtgtaagtgc caagaattat ttcagtttgt agttcccttg taagcagggt
     3061 tagtgataca tttgtattta agctttctgg gctgatcatt tggaaatgtc tcagtccagt
     3121 acctattgaa tgttatttgc ttaacctgaa gctaaataaa acttgttaac tacacccatt
     3181 aattgataaa ttcaaagcac gttttttctg tttggtgttt ggtgtggtaa caattctgtg
     3241 tatgtgtgtt ttatttagct tcggttaagt agcataacaa cctccaagca ctgaactttt
     3301 tttaataggt aatttaaact ttgcctatcg gcaaaatttt caatcaattg tacgccaaag
     3361 tgttgcatga tcaacgtttg acttattttt gtatttacta aatactgaat ttcgccgtga
     3421 cgctttttac agatggaaat tcacggcaaa atgttttttg ctaactttgc tatgtaaaac
     3481 aagaaacttg gcactcggtt attactaaat aaactggtaa aaaataacca ttagaaccaa
     3541 aaagaacgaa aaccagtaca cccttgccag ttttcaagct tttgctatga cgactctaat
     3601 aatcgggttt aacaccattc cgctttgaga aaattatcct tgtacagcaa gtaacagtca
     3661 atgctaaacg caccgctaca aatccttaag tttttccagt agcgatttac cttcttggta
     3721 actgcccgcc ttgatagccc agaatgtctc aattgatccc cgggcccggg cccgacgtcc
     3781 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     3841 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3901 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3961 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4021 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4081 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4321 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4681 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4801 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4921 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4981 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5041 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5101 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5161 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5221 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5281 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5341 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5401 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5461 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5521 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5581 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5641 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5701 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5761 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5821 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6001 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6061 c
//
