LOCUS       40924_49242        6852 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD052, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  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 6852)
  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 6852)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6852)
  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..6852
                     /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="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    429..434
                     /label=SacI
                     /note="SacI"
     CDS             complement(492..665)
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     regulatory      complement(667..789)
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
                     /regulatory_class="promoter"
     regulatory      complement(684..704)
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen
                     pTrcHis2"
                     /regulatory_class="other"
     protein_bind    686..702
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(710..740)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    755..776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    790..795
                     /label=SacI
                     /note="SacI"
     regulatory      796..1445
                     /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    1195..1200
                     /label=MfeI
                     /note="MfeI"
     CDS             1446..2567
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDU1; derived from pAM0505; Nostoc sp. PCC7524"
                     /protein_id="AII99679.1"
                     /translation="MISPESLNTNVSLTLLTRELQTEIILGTYTVRVHTRIGREPCARL
                     WYLCRALDKDGSGHLTLPLPVVQTFLDCSDKSVYRWLQDGKKIGAFRRYKIKAGMITVY
                     LGGMFQVCYNLNLKRWGDVAVVPLVQVLSDLRSLTTGIVTQSFQQKSRYAANRQLKPEY
                     RKLFGAPHPNELVKDTRQSSLKSPEGEVPCVLHISSSRIFVSKSFIHYGTSQKAVSCEL
                     GIHKRTVRRHQKQLGMNRRQLCQAKIEYNQLRHARNNDASEFWAFTGTKTDIGYQVMGD
                     AVVFSDGIAPGAKKRQPNTYQIDATEFDGRLFKVGDKVFMNRCNIYREQFTLTTMSAAR
                     RKYHFKLSQCHFSENRAGRVGNRFVIGCHSGEI"
     misc_feature    2262..2267
                     /label=EcoRV
                     /note="EcoRV"
     regulatory      2646..2673
                     /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    3084..3089
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(3188..3739)
                     /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="AII99678.1"
                     /translation="MKVNGNGRAKILTSDELRRLFSDGFTTPRDRVLFGICLFTGCRVS
                     EALALQTTDIKGETLTFRKSTTKGKLKTRVVDIQPGLAALMADYHPKPGTLFPGMRGVS
                     DRLTRYAADKILRDAAKRIGLEGISTHSFRRTALNQMSSAGIPLRHIQEISGHNDLGTL
                     QRYLEVTPEQRRKAVSVIGF"
     misc_feature    3276..3281
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    3617..3622
                     /label=AgeI
                     /note="AgeI"
     misc_feature    4044..4049
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(4111..4464)
                     /codon_start=1
                     /product="hyp1"
                     /label=hyp1
                     /note="hypothetical protein; derived from pAM505; Nostoc
                     sp. PCC7524"
                     /protein_id="AII99677.1"
                     /translation="MADKTLATFRIDSEEWESFKNLASSESSNASALLTEFVRWYLAGN
                     RFNTPTSHTPTHLDTSLEQRIDNIEQRLDKVTTNNLDNIDEFIDKRIEDNLATRLDKLQ
                     SQLEELRGKSKAR"
     misc_feature    4163..4168
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4220..4225
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4244..4249
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4289..4294
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4533..4538
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4537..4542
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    4541..4546
                     /label=XbaI
                     /note="XbaI"
     misc_feature    complement(4547..4567)
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    4565..4570
                     /label=ZraI
                     /note="ZraI"
     misc_feature    4589..4594
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(4631..4647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4655..4671)
                     /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(4679..4709)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4724..4745)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5033..5621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5795..6652)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6653..6757)
                     /label=AmpR promoter
     misc_feature    6783..6788
                     /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 gctcctactc cagccagctt tccggcaccg cttctggtgc cggaaaccag
      481 gcaaagcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      541 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      601 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatc cgtaatcatg
      661 gtcattagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      781 cgttgcgctg agctcagaca ttctgggcta tcaaggcggg cagttaccaa gaaggtaaat
      841 cgctactgga aaaacttaag gatttgtagc ggtgcgttta gcattgactg ttacttgctg
      901 tacaaggata attttctcaa agcggaatgg tgttaaaccc gattattaga gtcgtcatag
      961 caaaagcttg aaaactggca agggtgtact ggttttcgtt ctttttggtt ctaatggtta
     1021 ttttttacca gtttatttag taataaccga gtgccaagtt tcttgtttta catagcaaag
     1081 ttagcaaaaa acattttgcc gtgaatttcc atctgtaaaa agcgtcacgg cgaaattcag
     1141 tatttagtaa atacaaaaat aagtcaaacg ttgatcatgc aacactttgg cgtacaattg
     1201 attgaaaatt ttgccgatag gcaaagttta aattacctat taaaaaaagt tcagtgcttg
     1261 gaggttgtta tgctacttaa ccgaagctaa ataaaacaca catacacaga attgttacca
     1321 caccaaacac caaacagaaa aaacgtgctt tgaatttatc aattaatggg tgtagttaac
     1381 aagttttatt tagcttcagg ttaagcaaat aacattcaat aggtactgga ctgagacatt
     1441 tccaaatgat cagcccagaa agcttaaata caaatgtatc actaaccctg cttacaaggg
     1501 aactacaaac tgaaataatt cttggcactt acacggtcag ggtacacact cgcattggtc
     1561 gagaaccttg tgcgcggttg tggtatttgt gtcgagcgct tgataaggat gggtctggtc
     1621 atttaacatt gccattgcct gtagtacaga cgttcttaga ttgtagtgat aagtctgttt
     1681 accgttggtt acaggatgga aaaaaaatag gagcattccg ccgctacaaa attaaagcag
     1741 gaatgataac cgtctacttg ggcggtatgt ttcaagtttg ttacaaccta aatttaaaaa
     1801 gatggggtga tgttgcagta gtgcctttag tccaagtgtt aagcgattta cgctcactga
     1861 ccactggcat agtcacacaa agctttcaac aaaagagccg ctatgcggcc aacagacaac
     1921 ttaaaccaga gtacagaaaa ctttttggag cgccacaccc gaacgagcta gtgaaggaca
     1981 caaggcaatc ctctcttaaa tcgccagagg gggaagtacc ttgtgtactt cacattagtt
     2041 cgtcgcggat attcgtctca aaaagtttca tccactatgg tacatctcaa aaagctgtca
     2101 gctgtgagtt gggtattcac aagcgcacag tgcgacgaca tcaaaagcag ctgggcatga
     2161 atcgccgtca actttgccaa gccaaaattg agtacaatca actccggcac gctcggaata
     2221 atgacgcgtc tgaattttgg gctttcactg gaaccaaaac tgatatcggt tatcaagtta
     2281 tgggtgacgc ggttgtgttt tctgatggca ttgcccctgg agccaaaaag agacagccca
     2341 acacttacca aattgacgcg actgaatttg acggtcgatt attcaaagtg ggtgacaagg
     2401 tgttcatgaa tcggtgcaac atctaccgcg aacaattcac cctaacaacg atgtcagcag
     2461 cacggcggaa gtaccatttc aaactgtcac agtgccactt cagcgaaaat cgagccggtc
     2521 gggtaggcaa ccgttttgta ataggttgcc actctggcga gatatgaagg acatttttaa
     2581 gaattttttt ttagcgtgaa atgctgcggc aaccagtata gtgatcgaca cgaactagtg
     2641 cagcatgacc gagagcctat agggtcgcgg ctttttggca atagagatcg tatgcagatc
     2701 atgcgtagta ctttcttcga ccttaacccc aaagaaagct tattagatgg agaaggtcac
     2761 aattagaaaa tattcatcgt cccgattgac ccttaacctt taatcggttt ccacttgtaa
     2821 aaaattcggc gagccattct tcttaaccct cagagtcgaa cttcatacaa gaatatacaa
     2881 accctttggg tttaccacta aaacaacggc gataaaaacc taataggttt ccacagaaac
     2941 gtctaaggat ttaggcgtgt tgttttgggg caatttgaga aaatattaaa ccagtcacaa
     3001 gttaacttgt cgaagaaata ttgcagcatt ctgcaacaca gggaaaggta taacgtttcc
     3061 aaagtccgga acgtgtgcag agttctagaa ccgatggata ctttaaaagt atcctcaagt
     3121 gtagagcgtg tgtttaggct gcgtcttttt ttagcaccca taggggtcta aacagcgttg
     3181 gcgtacatta gaagccaatc acggatacag ctttgcgtcg ctgttcgggt gtaacttcaa
     3241 gatagcgttg cagtgtgcca aggtcattgt gaccggatat ctcttgaatg tgtcgcaacg
     3301 ggataccggc gctagacatt tggttgaggg cagtacggcg gaaactgtgg gtactgatgc
     3361 cttctagccc gattcttttg gctgcatcgc gcaagatttt atccgccgcg tatcgcgtga
     3421 gcctatcgct gactcccctc atgccaggga acagggttcc cggtttgggg tgatagtcag
     3481 ccatgagtgc ggctagtcct ggctggatgt caaccacgcg ggttttgagt ttccctttgg
     3541 tggtagactt cctaaaggtt agtgtttcgc ctttaatgtc cgttgtttgg agtgctagag
     3601 cttcactaac gcggcaaccg gtgaatagac agatgccaaa caaaacgcga tcgcgcggtg
     3661 tggtgaatcc gtcgctaaac agtcgcctga gttcgtcgga ggtgagtatt ttggctcgtc
     3721 cgttgccgtt aaccttcata acccttacag attaagcatt ccaatactct acacgttatc
     3781 tgcattttgt ggaattggtg gctatatcta ttgattcttc gtatgctttc tgttgaaagc
     3841 cagtgctagt ttagccaagc ttatcaatcg ttgcacggta aaatcaggct ttcgacgtaa
     3901 ggacagaatt ttgataactt cagcttatgt tgtctcctga agaaagcaac tctttgataa
     3961 acgcgtcgat ggctttcccg gcggctgact gcctacccat ttttagctta ttgagggtgc
     4021 gatcgcgggt tgcctccaat ttgtctagat tgtctatact attgtctata ttctttctgt
     4081 cttgcccggt agcttgtcct tctgcctgaa ctaccgggct ttcgattttc cccgcagttc
     4141 ctccagttgc gattgaagtt tgtctagacg tgttgctaga ttatcttcta tacgcttgtc
     4201 tataaattca tctatattgt ctagattatt agttgtgact ttatctagac gttgttcaat
     4261 attgtctata cgctgttcga gggatgtgtc tagatgggtg ggagtgtgag aagtgggagt
     4321 attaaacctg ttacctgcca aataccaacg aacaaattct gttaacagtg ctgaggcgtt
     4381 ggaactttca gaactagcaa ggtttttaaa agactcccat tcttcggagt caatacgaaa
     4441 agtggctaga gttttgtctg ccatgattag ataatgtcta tacttgtata gatagcagta
     4501 taaactattg tctatattag tgtctatata agtctagata tctagaatcc ccccgggggc
     4561 ccccgacgtc ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg
     4621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4681 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4741 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5641 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6781 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6841 ggccctttcg tc
//
