LOCUS       40924_49332        6221 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD077, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6221)
  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 6221)
  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 6221)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6221)
  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..6221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..6
                     /label=ZraI
                     /note="ZraI"
     misc_feature    4..24
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    25..30
                     /label=XbaI
                     /note="XbaI"
     misc_recomb     31..1091
                     /note="A7120NS1-RA; right arm of the homologous
                     recombination site NS1 (alr3858_D748-alr3858_D749) into the
                     chromosome of Anabaena sp. PCC7120 derived from INSD 
                     accession NC_003272"
     misc_feature    31..38
                     /label=SwaI
                     /note="SwaI"
     misc_feature    515..520
                     /label=XbaI
                     /note="XbaI"
     rep_origin      1128..1716
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1902..2042)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    2205..2210
                     /label=EcoRV
                     /note="EcoRV"
     CDS             complement(2233..3651)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     regulatory      complement(3652..3957)
                     /note="PsacB; derived from Bacillus subtilis INSD accession
                     AAA72307"
                     /regulatory_class="promoter"
     misc_recomb     3958..4965
                     /note="A7120NS1-LA; right arm of the homologous
                     recombination site NS1 (alr3858_D748-alr3858_D749) into the
                     chromosome of Anabaena sp. PCC7120 derived from INSD 
                     accession NC_003272"
     misc_feature    4697..4702
                     /label=MfeI
                     /note="MfeI"
     misc_feature    4966..4971
                     /label=NheI
                     /note="NheI"
     misc_feature    4972..4992
                     /note="GC; GC-adaptor for seamless assembly, flanking BHR 
                     replicons or neutral sites; synthetic"
     misc_feature    4990..4995
                     /label=ZraI
                     /note="ZraI"
     CDS             complement(5119..5976)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5977..6081)
                     /label=AmpR promoter
     misc_feature    6107..6112
                     /label=ZraI
                     /note="ZraI"
ORIGIN
        1 gacgtcgggg gcccccgggg ggattctaga atttaaatgg gatgccttat ggaagtctaa
       61 ttgtatggtt agagttgtaa ttatccccaa ttttgaaacc tatcaactat attattaagg
      121 cagtgaagac ttgccataac ctatatttct cgcatgaagt aatctgctga gaactatgag
      181 ttgtgtaata tattgtattt ttattaagtg taggtttaat tgcatagagt tagccgttag
      241 aactaattat cccagaatag atacctcaaa ttctcagtag cagaaattaa atagataaca
      301 aaatatacaa aatgccttca gtatttaaaa cttcttctca gaagacaaac acagttagta
      361 accagatgaa taactcatca aaatcatcaa cagaagtatt agctgtgtct cctgcatccc
      421 aaaggtatac tatgcaaaaa ttgccatgta gagacaatat taataagttg tggaagttat
      481 acagatcaat tttaagtaaa tgtatgtata cttctctaga attagctgtg cattcaccta
      541 tggtgagcca aacactatct cctgggtatg ctcatccaac atctaaggta aggcgaagcc
      601 cagtccaaat gttggtgttt ggtctcatag cagactttca cacttacaat cttgttaagg
      661 ttggcagtag tgataacaat caagaagagt gccaaaattt ttagtgctgt caaacttaat
      721 agttgactcc tagatgtgta acacctaaaa aagtcgttta tctaccttcc cgaatccaat
      781 gacacttgat aataaccaag agcttaccta tagaaattcc caatcgttgg gacagccagg
      841 tttctcactg gcagttaact caagcaaccc tttcaatcac tctgggctga attttggaca
      901 aaataacgac agtaagaaga tttctgtgga aaataaccgt attggcgaga ttgttcctgg
      961 gcgcgttgcc aatatcaaag taattggtgt tggcggtggt ggtggtaatg ctgttaaccg
     1021 catgattgaa tctgatgtct ctggtgtaga gttttggtca attaatactg atgctcaagc
     1081 tttgaccttg gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1261 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1321 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1381 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1441 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1501 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1561 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1621 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1681 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1741 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1801 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1861 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     1921 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     1981 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2041 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2101 ccgcttacag acaagctccg cccggttagc catttgcctg cttttatata gttcatatgg
     2161 gattcacctt tatgttgata agaaataaaa gaaaatgcca ataggatatc ggcattttct
     2221 tttgcgtttt tatttgttaa ctgttaattg tccttgttca aggatgctgt ctttgacaac
     2281 agatgttttc ttgcctttga tgttcagcag gaagcttggc gcaaacgttg attgtttgtc
     2341 tgcgtagaat cctctgtttg tcatatagct tgtaatcacg acattgtttc ctttcgcttg
     2401 aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat ccatttttaa
     2461 cacaaggcca gttttgttca gcggcttgta tgggccagtt aaagaattag aaacataacc
     2521 aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc atttttgatc cgcgggagtc
     2581 agtgaacagg taccatttgc cgttcatttt aaagacgttc gcgcgttcaa tttcatctgt
     2641 tactgtgtta gatgcaatca gcggtttcat cacttttttc agtgtgtaat catcgtttag
     2701 ctcaatcata ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat cgctttgcag
     2761 aagtttttga ctttcttgac ggaagaatga tgtgcttttg ccatagtatg ctttgttaaa
     2821 taaagattct tcgccttggt agccatcttc agttccagtg tttgcttcaa atactaagta
     2881 tttgtggcct ttatcttcta cgtagtgagg atctctcagc gtatggttgt cgcctgagct
     2941 gtagttgcct tcatcgatga actgctgtac attttgatac gtttttccgt caccgtcaaa
     3001 gattgattta taatcctcta caccgttgat gttcaaagag ctgtctgatg ctgatacgtt
     3061 aacttgtgca gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat cagtgtagaa
     3121 taaacggatt tttccgtcag atgtaaatgt ggctgaacct gaccattctt gtgtttggtc
     3181 ttttaggata gaatcatttg catcgaattt gtcgctgtct ttaaagacgc ggccagcgtt
     3241 tttccagctg tcaatagaag tttcgccgac tttttgatag aacatgtaaa tcgatgtgtc
     3301 atccgcattt ttaggatctc cggctaatgc aaagacgatg tggtagccgt gatagtttgc
     3361 gacagtgccg tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc cttttgcaga
     3421 agagatattt ttaattgtgg acgaatcgaa ttcaggaact tgatattttt catttttttg
     3481 ctgttcaggg atttgcagca tatcatggcg tgtaatatgg gaaatgccgt atgtttcctt
     3541 atatggcttt tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg ccagcagtgc
     3601 ggtagtaaag gttaatactg ttgcttgttt tgcaaacttt ttgatgttca tcgttcatgt
     3661 ctcctttttt atgtactgtg ttagcggtct gcttcttcca gccctcctgt ttgaagatgg
     3721 caagttagtt acgcacaata aaaaaagacc taaaatatgt aaggggtgac gccaaagtat
     3781 acactttgcc ctttacacat tttaggtctt gcctgcttta tcagtaacaa acccgcgcga
     3841 tttacttttc gacctcattc tattagactc tcgtttggat tgcaactggt ctattttcct
     3901 cttttgtttg atagaaaatc ataaaaggat ttgcagacta cgggcctaaa gaactagcga
     3961 tcgctgactc tttgaagaaa caaccaacca ttgcccaagc aactgtcaat cgtcgtcttt
     4021 ttccaccagg tttaattatc gaaatcgaag aaagaatacc tgtcgcagtg gctcaaagga
     4081 ggagagaaca atctaatagt actagtaaca aacaaacaca cacaggttta atcgatgcca
     4141 atggagtttg gattcctctg gaaaaatata cactagtaaa tcctcagttc aaactcccta
     4201 cacttaaggt aattggatta ccagaacaat atgctcccta ctggagtaaa ctatatccct
     4261 atatcagcca aagttccata aaaattacgg aaattgatta ccaagaccca aataacttaa
     4321 ttttgaaaac tgaattagga acagtttatc ttggtgctac aagcgcccaa ttagctgacc
     4381 agattaattt actagcacaa ttacgccaca tcaatacaaa actaaatcct tcggagatag
     4441 actatattga tttaaaaaat cctgaatcgc ctttagtaca tatgatccaa aataaagaga
     4501 caattaaaaa tcaaacgcca taatcatgtg tcaaaaattc cttatatatt aatgaatttt
     4561 attttataaa acaatattat tgtttactct acgctttaca tagaaattgt gaaaagctct
     4621 tttccagacg cttagaactt agggacaaaa atgatttctg gatacttgcg ctcaccgtgt
     4681 agtacgtttc cagtggcaat tgttcagtct gtagcagata ttcaaactga ttgactttac
     4741 gactgaacag cttgataggc ttagatagta ttgctattaa gtttcagttt ggtcacaaaa
     4801 agtaacaaat taattaatca gtcagatttt tattcctttg ttatctttaa attttcgcat
     4861 caataaataa cacagatagg agttgacttt gatttctgtt ggcgcagtgt ggcatagcta
     4921 taggtacttg tgtatctagg ggatggtcac tccttcacca acaaagctag catcgcgcgc
     4981 gcgcgcgcgg acgtcctcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5041 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5101 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5161 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5221 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5281 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5341 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5401 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5461 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5521 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5581 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5641 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5701 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5761 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5821 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5881 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5941 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6001 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6061 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6121 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6181 gcgtttcggt gatgacggtg aaaacctctg acacatgcag c
//
