LOCUS       40924_49252        4660 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD054, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4660)
  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 4660)
  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 4660)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4660)
  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..4660
                     /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..2375
                     /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"
     regulatory      complement(471..498)
                     /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(577..1698)
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDU1; derived from pAM0505; Nostoc sp. PCC7524"
                     /protein_id="AII99685.1"
                     /translation="MISPESLNTNVSLTLLTRELQTEIILGTYTVRVHTRIGREPCARL
                     WYLCRALDKDGSGHLTLPLPVVQTFLDCSDKSVYRWLQDGKKIGAFRRYKIKAGMITVY
                     LGGMFQVCYNLNLKRWGDVAVVPLVQVLSDLRSLTTGIVTQSFQQKSRYAANRQLKPEY
                     RKLFGAPHPNELVKDTRQSSLKSPEGEVPCVLHISSSRIFVSKSFIHYGTSQKAVSCEL
                     GIHKRTVRRHQKQLGMNRRQLCQAKIEYNQLRHARNNDASEFWAFTGTKTDIGYQVMGD
                     AVVFSDGIAPGAKKRQPNTYQIDATEFDGRLFKVGDKVFMNRCNIYREQFTLTTMSAAR
                     RKYHFKLSQCHFSENRAGRVGNRFVIGCHSGEI"
     misc_feature    877..882
                     /label=EcoRV
                     /note="EcoRV"
     regulatory      complement(1699..2348)
                     /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    1944..1949
                     /label=MfeI
                     /note="MfeI"
     misc_feature    2349..2354
                     /label=MfeI
                     /note="MfeI"
     misc_feature    2355..2375
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    2373..2378
                     /label=ZraI
                     /note="ZraI"
     misc_feature    2397..2402
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2439..2455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2463..2479)
                     /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(2487..2517)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2532..2553)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2841..3429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3603..4460)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4461..4565)
                     /label=AmpR promoter
     misc_feature    4591..4596
                     /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 tagaactacg catgatctgc atacgatctc tattgccaaa aagccgcgac
      481 cctataggct ctcggtcatg ctgcactagt tcgtgtcgat cactatactg gttgccgcag
      541 catttcacgc taaaaaaaaa ttcttaaaaa tgtccttcat atctcgccag agtggcaacc
      601 tattacaaaa cggttgccta cccgaccggc tcgattttcg ctgaagtggc actgtgacag
      661 tttgaaatgg tacttccgcc gtgctgctga catcgttgtt agggtgaatt gttcgcggta
      721 gatgttgcac cgattcatga acaccttgtc acccactttg aataatcgac cgtcaaattc
      781 agtcgcgtca atttggtaag tgttgggctg tctctttttg gctccagggg caatgccatc
      841 agaaaacaca accgcgtcac ccataacttg ataaccgata tcagttttgg ttccagtgaa
      901 agcccaaaat tcagacgcgt cattattccg agcgtgccgg agttgattgt actcaatttt
      961 ggcttggcaa agttgacggc gattcatgcc cagctgcttt tgatgtcgtc gcactgtgcg
     1021 cttgtgaata cccaactcac agctgacagc tttttgagat gtaccatagt ggatgaaact
     1081 ttttgagacg aatatccgcg acgaactaat gtgaagtaca caaggtactt ccccctctgg
     1141 cgatttaaga gaggattgcc ttgtgtcctt cactagctcg ttcgggtgtg gcgctccaaa
     1201 aagttttctg tactctggtt taagttgtct gttggccgca tagcggctct tttgttgaaa
     1261 gctttgtgtg actatgccag tggtcagtga gcgtaaatcg cttaacactt ggactaaagg
     1321 cactactgca acatcacccc atctttttaa atttaggttg taacaaactt gaaacatacc
     1381 gcccaagtag acggttatca ttcctgcttt aattttgtag cggcggaatg ctcctatttt
     1441 ttttccatcc tgtaaccaac ggtaaacaga cttatcacta caatctaaga acgtctgtac
     1501 tacaggcaat ggcaatgtta aatgaccaga cccatcctta tcaagcgctc gacacaaata
     1561 ccacaaccgc gcacaaggtt ctcgaccaat gcgagtgtgt accctgaccg tgtaagtgcc
     1621 aagaattatt tcagtttgta gttcccttgt aagcagggtt agtgatacat ttgtatttaa
     1681 gctttctggg ctgatcattt ggaaatgtct cagtccagta cctattgaat gttatttgct
     1741 taacctgaag ctaaataaaa cttgttaact acacccatta attgataaat tcaaagcacg
     1801 ttttttctgt ttggtgtttg gtgtggtaac aattctgtgt atgtgtgttt tatttagctt
     1861 cggttaagta gcataacaac ctccaagcac tgaacttttt ttaataggta atttaaactt
     1921 tgcctatcgg caaaattttc aatcaattgt acgccaaagt gttgcatgat caacgtttga
     1981 cttatttttg tatttactaa atactgaatt tcgccgtgac gctttttaca gatggaaatt
     2041 cacggcaaaa tgttttttgc taactttgct atgtaaaaca agaaacttgg cactcggtta
     2101 ttactaaata aactggtaaa aaataaccat tagaaccaaa aagaacgaaa accagtacac
     2161 ccttgccagt tttcaagctt ttgctatgac gactctaata atcgggttta acaccattcc
     2221 gctttgagaa aattatcctt gtacagcaag taacagtcaa tgctaaacgc accgctacaa
     2281 atccttaagt ttttccagta gcgatttacc ttcttggtaa ctgcccgcct tgatagccca
     2341 gaatgtctca attgatcccc gggcccgggc ccgacgtcct cggtacccgg ggatcctcta
     2401 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     2461 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2521 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2581 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2641 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2701 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2761 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3001 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3481 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3541 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3601 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3661 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3721 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3781 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3841 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3901 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3961 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4021 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4081 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4141 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4201 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4261 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4321 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4381 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4441 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4501 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4561 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4621 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
