LOCUS       40924_49152        5612 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD028, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  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 5612)
  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 5612)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5612)
  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..5612
                     /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..428)
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     Escherichia coli origins for KO; synthetic"
     misc_feature    429..434
                     /label=SacI
                     /note="SacI"
     misc_feature    517..522
                     /label=EcoRV
                     /note="EcoRV"
     CDS             complement(545..1963)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     regulatory      complement(1964..2269)
                     /note="PsacB; derived from Bacillus subtilis INSD accession
                     AAA72307"
                     /regulatory_class="promoter"
     misc_feature    2269..2274
                     /label=SacI
                     /note="SacI"
     misc_feature    2350..2490
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2676..3264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3301..3306
                     /label=MfeI
                     /note="MfeI"
     misc_feature    complement(3307..3327)
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     Escherichia coli origins for KO; synthetic"
     misc_feature    3325..3330
                     /label=ZraI
                     /note="ZraI"
     misc_feature    3349..3354
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(3391..3407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3415..3431)
                     /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(3439..3469)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3484..3505)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3734..4407)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(3793..4381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4555..5412)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5413..5517)
                     /label=AmpR promoter
     misc_feature    5543..5548
                     /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 cccccggggg
      421 cccccgatga gctccggtta gccatttgcc tgcttttata tagttcatat gggattcacc
      481 tttatgttga taagaaataa aagaaaatgc caataggata tcggcatttt cttttgcgtt
      541 tttatttgtt aactgttaat tgtccttgtt caaggatgct gtctttgaca acagatgttt
      601 tcttgccttt gatgttcagc aggaagcttg gcgcaaacgt tgattgtttg tctgcgtaga
      661 atcctctgtt tgtcatatag cttgtaatca cgacattgtt tcctttcgct tgaggtacag
      721 cgaagtgtga gtaagtaaag gttacatcgt taggatcaag atccattttt aacacaaggc
      781 cagttttgtt cagcggcttg tatgggccag ttaaagaatt agaaacataa ccaagcatgt
      841 aaatatcgtt agacgtaatg ccgtcaatcg tcatttttga tccgcgggag tcagtgaaca
      901 ggtaccattt gccgttcatt ttaaagacgt tcgcgcgttc aatttcatct gttactgtgt
      961 tagatgcaat cagcggtttc atcacttttt tcagtgtgta atcatcgttt agctcaatca
     1021 taccgagagc gccgtttgct aactcagccg tgcgtttttt atcgctttgc agaagttttt
     1081 gactttcttg acggaagaat gatgtgcttt tgccatagta tgctttgtta aataaagatt
     1141 cttcgccttg gtagccatct tcagttccag tgtttgcttc aaatactaag tatttgtggc
     1201 ctttatcttc tacgtagtga ggatctctca gcgtatggtt gtcgcctgag ctgtagttgc
     1261 cttcatcgat gaactgctgt acattttgat acgtttttcc gtcaccgtca aagattgatt
     1321 tataatcctc tacaccgttg atgttcaaag agctgtctga tgctgatacg ttaacttgtg
     1381 cagttgtcag tgtttgtttg ccgtaatgtt taccggagaa atcagtgtag aataaacgga
     1441 tttttccgtc agatgtaaat gtggctgaac ctgaccattc ttgtgtttgg tcttttagga
     1501 tagaatcatt tgcatcgaat ttgtcgctgt ctttaaagac gcggccagcg tttttccagc
     1561 tgtcaataga agtttcgccg actttttgat agaacatgta aatcgatgtg tcatccgcat
     1621 ttttaggatc tccggctaat gcaaagacga tgtggtagcc gtgatagttt gcgacagtgc
     1681 cgtcagcgtt ttgtaatggc cagctgtccc aaacgtccag gccttttgca gaagagatat
     1741 ttttaattgt ggacgaatcg aattcaggaa cttgatattt ttcatttttt tgctgttcag
     1801 ggatttgcag catatcatgg cgtgtaatat gggaaatgcc gtatgtttcc ttatatggct
     1861 tttggttcgt ttctttcgca aacgcttgag ttgcgcctcc tgccagcagt gcggtagtaa
     1921 aggttaatac tgttgcttgt tttgcaaact ttttgatgtt catcgttcat gtctcctttt
     1981 ttatgtactg tgttagcggt ctgcttcttc cagccctcct gtttgaagat ggcaagttag
     2041 ttacgcacaa taaaaaaaga cctaaaatat gtaaggggtg acgccaaagt atacactttg
     2101 ccctttacac attttaggtc ttgcctgctt tatcagtaac aaacccgcgc gatttacttt
     2161 tcgacctcat tctattagac tctcgtttgg attgcaactg gtctattttc ctcttttgtt
     2221 tgatagaaaa tcataaaagg atttgcagac tacgggccta aagaactaga gctcagcttg
     2281 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2341 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     2401 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     2461 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     2521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3301 caattgatcg ggcccgggcc cggggacgtc ctcggtaccc ggggatcctc tagagtcgac
     3361 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3421 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3481 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3541 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3601 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3661 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3721 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3781 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3841 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3901 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3961 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4021 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4081 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4141 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4201 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4261 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4321 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4441 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4681 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5521 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5581 taaaaatagg cgtatcacga ggccctttcg tc
//
