LOCUS       40924_49237        6491 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD051, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6491)
  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 6491)
  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 6491)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6491)
  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..6491
                     /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    complement(408..4206)
                     /note="CVD-CYA; device for the GC-adaptor assembly: 
                     cyanobacterial replicons"
     misc_feature    complement(408..428)
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    429..434
                     /label=SacI
                     /note="SacI"
     regulatory      435..1084
                     /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    834..839
                     /label=MfeI
                     /note="MfeI"
     CDS             1085..2206
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDU1; derived from pAM0505; Nostoc sp. PCC7524"
                     /protein_id="AII99675.1"
                     /translation="MISPESLNTNVSLTLLTRELQTEIILGTYTVRVHTRIGREPCARL
                     WYLCRALDKDGSGHLTLPLPVVQTFLDCSDKSVYRWLQDGKKIGAFRRYKIKAGMITVY
                     LGGMFQVCYNLNLKRWGDVAVVPLVQVLSDLRSLTTGIVTQSFQQKSRYAANRQLKPEY
                     RKLFGAPHPNELVKDTRQSSLKSPEGEVPCVLHISSSRIFVSKSFIHYGTSQKAVSCEL
                     GIHKRTVRRHQKQLGMNRRQLCQAKIEYNQLRHARNNDASEFWAFTGTKTDIGYQVMGD
                     AVVFSDGIAPGAKKRQPNTYQIDATEFDGRLFKVGDKVFMNRCNIYREQFTLTTMSAAR
                     RKYHFKLSQCHFSENRAGRVGNRFVIGCHSGEI"
     misc_feature    1901..1906
                     /label=EcoRV
                     /note="EcoRV"
     regulatory      2285..2312
                     /note="rho-ITTS; rho-ITTS; rho-independent transcriptional 
                     termination signal, downstream of repA in pDU1, from 
                     pAM505; Nostoc sp. PCC7524"
                     /regulatory_class="terminator"
     misc_feature    2723..2728
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(2827..3378)
                     /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="AII99674.1"
                     /translation="MKVNGNGRAKILTSDELRRLFSDGFTTPRDRVLFGICLFTGCRVS
                     EALALQTTDIKGETLTFRKSTTKGKLKTRVVDIQPGLAALMADYHPKPGTLFPGMRGVS
                     DRLTRYAADKILRDAAKRIGLEGISTHSFRRTALNQMSSAGIPLRHIQEISGHNDLGTL
                     QRYLEVTPEQRRKAVSVIGF"
     misc_feature    2915..2920
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    3256..3261
                     /label=AgeI
                     /note="AgeI"
     misc_feature    3683..3688
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(3750..4103)
                     /codon_start=1
                     /product="hyp1"
                     /label=hyp1
                     /note="hypothetical protein; derived from pAM505; Nostoc
                     sp. PCC7524"
                     /protein_id="AII99673.1"
                     /translation="MADKTLATFRIDSEEWESFKNLASSESSNASALLTEFVRWYLAGN
                     RFNTPTSHTPTHLDTSLEQRIDNIEQRLDKVTTNNLDNIDEFIDKRIEDNLATRLDKLQ
                     SQLEELRGKSKAR"
     misc_feature    3802..3807
                     /label=XbaI
                     /note="XbaI"
     misc_feature    3859..3864
                     /label=XbaI
                     /note="XbaI"
     misc_feature    3883..3888
                     /label=XbaI
                     /note="XbaI"
     misc_feature    3928..3933
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4172..4177
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4176..4181
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    4180..4185
                     /label=XbaI
                     /note="XbaI"
     misc_feature    complement(4186..4206)
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    4204..4209
                     /label=ZraI
                     /note="ZraI"
     misc_feature    4228..4233
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(4270..4286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4294..4310)
                     /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(4318..4348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4363..4384)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4672..5260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5434..6291)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6292..6396)
                     /label=AmpR promoter
     misc_feature    6422..6427
                     /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 gggcccgggc
      421 ccggggatga gctcagacat tctgggctat caaggcgggc agttaccaag aaggtaaatc
      481 gctactggaa aaacttaagg atttgtagcg gtgcgtttag cattgactgt tacttgctgt
      541 acaaggataa ttttctcaaa gcggaatggt gttaaacccg attattagag tcgtcatagc
      601 aaaagcttga aaactggcaa gggtgtactg gttttcgttc tttttggttc taatggttat
      661 tttttaccag tttatttagt aataaccgag tgccaagttt cttgttttac atagcaaagt
      721 tagcaaaaaa cattttgccg tgaatttcca tctgtaaaaa gcgtcacggc gaaattcagt
      781 atttagtaaa tacaaaaata agtcaaacgt tgatcatgca acactttggc gtacaattga
      841 ttgaaaattt tgccgatagg caaagtttaa attacctatt aaaaaaagtt cagtgcttgg
      901 aggttgttat gctacttaac cgaagctaaa taaaacacac atacacagaa ttgttaccac
      961 accaaacacc aaacagaaaa aacgtgcttt gaatttatca attaatgggt gtagttaaca
     1021 agttttattt agcttcaggt taagcaaata acattcaata ggtactggac tgagacattt
     1081 ccaaatgatc agcccagaaa gcttaaatac aaatgtatca ctaaccctgc ttacaaggga
     1141 actacaaact gaaataattc ttggcactta cacggtcagg gtacacactc gcattggtcg
     1201 agaaccttgt gcgcggttgt ggtatttgtg tcgagcgctt gataaggatg ggtctggtca
     1261 tttaacattg ccattgcctg tagtacagac gttcttagat tgtagtgata agtctgttta
     1321 ccgttggtta caggatggaa aaaaaatagg agcattccgc cgctacaaaa ttaaagcagg
     1381 aatgataacc gtctacttgg gcggtatgtt tcaagtttgt tacaacctaa atttaaaaag
     1441 atggggtgat gttgcagtag tgcctttagt ccaagtgtta agcgatttac gctcactgac
     1501 cactggcata gtcacacaaa gctttcaaca aaagagccgc tatgcggcca acagacaact
     1561 taaaccagag tacagaaaac tttttggagc gccacacccg aacgagctag tgaaggacac
     1621 aaggcaatcc tctcttaaat cgccagaggg ggaagtacct tgtgtacttc acattagttc
     1681 gtcgcggata ttcgtctcaa aaagtttcat ccactatggt acatctcaaa aagctgtcag
     1741 ctgtgagttg ggtattcaca agcgcacagt gcgacgacat caaaagcagc tgggcatgaa
     1801 tcgccgtcaa ctttgccaag ccaaaattga gtacaatcaa ctccggcacg ctcggaataa
     1861 tgacgcgtct gaattttggg ctttcactgg aaccaaaact gatatcggtt atcaagttat
     1921 gggtgacgcg gttgtgtttt ctgatggcat tgcccctgga gccaaaaaga gacagcccaa
     1981 cacttaccaa attgacgcga ctgaatttga cggtcgatta ttcaaagtgg gtgacaaggt
     2041 gttcatgaat cggtgcaaca tctaccgcga acaattcacc ctaacaacga tgtcagcagc
     2101 acggcggaag taccatttca aactgtcaca gtgccacttc agcgaaaatc gagccggtcg
     2161 ggtaggcaac cgttttgtaa taggttgcca ctctggcgag atatgaagga catttttaag
     2221 aatttttttt tagcgtgaaa tgctgcggca accagtatag tgatcgacac gaactagtgc
     2281 agcatgaccg agagcctata gggtcgcggc tttttggcaa tagagatcgt atgcagatca
     2341 tgcgtagtac tttcttcgac cttaacccca aagaaagctt attagatgga gaaggtcaca
     2401 attagaaaat attcatcgtc ccgattgacc cttaaccttt aatcggtttc cacttgtaaa
     2461 aaattcggcg agccattctt cttaaccctc agagtcgaac ttcatacaag aatatacaaa
     2521 ccctttgggt ttaccactaa aacaacggcg ataaaaacct aataggtttc cacagaaacg
     2581 tctaaggatt taggcgtgtt gttttggggc aatttgagaa aatattaaac cagtcacaag
     2641 ttaacttgtc gaagaaatat tgcagcattc tgcaacacag ggaaaggtat aacgtttcca
     2701 aagtccggaa cgtgtgcaga gttctagaac cgatggatac tttaaaagta tcctcaagtg
     2761 tagagcgtgt gtttaggctg cgtctttttt tagcacccat aggggtctaa acagcgttgg
     2821 cgtacattag aagccaatca cggatacagc tttgcgtcgc tgttcgggtg taacttcaag
     2881 atagcgttgc agtgtgccaa ggtcattgtg accggatatc tcttgaatgt gtcgcaacgg
     2941 gataccggcg ctagacattt ggttgagggc agtacggcgg aaactgtggg tactgatgcc
     3001 ttctagcccg attcttttgg ctgcatcgcg caagatttta tccgccgcgt atcgcgtgag
     3061 cctatcgctg actcccctca tgccagggaa cagggttccc ggtttggggt gatagtcagc
     3121 catgagtgcg gctagtcctg gctggatgtc aaccacgcgg gttttgagtt tccctttggt
     3181 ggtagacttc ctaaaggtta gtgtttcgcc tttaatgtcc gttgtttgga gtgctagagc
     3241 ttcactaacg cggcaaccgg tgaatagaca gatgccaaac aaaacgcgat cgcgcggtgt
     3301 ggtgaatccg tcgctaaaca gtcgcctgag ttcgtcggag gtgagtattt tggctcgtcc
     3361 gttgccgtta accttcataa cccttacaga ttaagcattc caatactcta cacgttatct
     3421 gcattttgtg gaattggtgg ctatatctat tgattcttcg tatgctttct gttgaaagcc
     3481 agtgctagtt tagccaagct tatcaatcgt tgcacggtaa aatcaggctt tcgacgtaag
     3541 gacagaattt tgataacttc agcttatgtt gtctcctgaa gaaagcaact ctttgataaa
     3601 cgcgtcgatg gctttcccgg cggctgactg cctacccatt tttagcttat tgagggtgcg
     3661 atcgcgggtt gcctccaatt tgtctagatt gtctatacta ttgtctatat tctttctgtc
     3721 ttgcccggta gcttgtcctt ctgcctgaac taccgggctt tcgattttcc ccgcagttcc
     3781 tccagttgcg attgaagttt gtctagacgt gttgctagat tatcttctat acgcttgtct
     3841 ataaattcat ctatattgtc tagattatta gttgtgactt tatctagacg ttgttcaata
     3901 ttgtctatac gctgttcgag ggatgtgtct agatgggtgg gagtgtgaga agtgggagta
     3961 ttaaacctgt tacctgccaa ataccaacga acaaattctg ttaacagtgc tgaggcgttg
     4021 gaactttcag aactagcaag gtttttaaaa gactcccatt cttcggagtc aatacgaaaa
     4081 gtggctagag ttttgtctgc catgattaga taatgtctat acttgtatag atagcagtat
     4141 aaactattgt ctatattagt gtctatataa gtctagatat ctagaatccc cccgggggcc
     4201 cccgacgtcc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
     4261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4321 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5101 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6421 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6481 gccctttcgt c
//
