LOCUS       40924_49257        5021 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD055, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5021)
  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 5021)
  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 5021)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5021)
  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..5021
                     /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..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="AII99687.1"
                     /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=SacI
                     /note="SacI"
     protein_bind    2368..2389
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2404..2434
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2442..2458
                     /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)."
     CDS             2479..2652
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    2710..2715
                     /label=SacI
                     /note="SacI"
     misc_feature    2716..2736
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    2734..2739
                     /label=ZraI
                     /note="ZraI"
     misc_feature    2758..2763
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2800..2816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2824..2840)
                     /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(2848..2878)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2893..2914)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3202..3790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3964..4821)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4822..4926)
                     /label=AmpR promoter
     misc_feature    4952..4957
                     /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 gaatgtctga gctcagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     2401 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     2461 tcacacagga aacagctaat gaccatgatt acggattcac tggccgtcgt tttacaacgt
     2521 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2581 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     2641 ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg
     2701 ctggagtagg agctcatccc cgggcccggg cccgacgtcc tcggtacccg gggatcctct
     2761 agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2821 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2881 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2941 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3001 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3061 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3121 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3181 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3241 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3301 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3361 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3421 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3481 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3541 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3601 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3661 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3721 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3781 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3841 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3901 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3961 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4021 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4081 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4141 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4201 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4261 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4321 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4381 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4441 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4501 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4561 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4621 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4681 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4741 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4801 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4861 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4921 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4981 attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
