LOCUS       Exported                5227 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD057, complete sequence.
ACCESSION   KM017884
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  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 5227)
  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 5227)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5227
                     /organism="Broad host range vector vector pCVD057"
                     /mol_type="other DNA"
                     /note="pFDAZ is a replication device carrying a minimal 
                     fragment of pFDA (an endogenous plasmid of Fremyella 
                     diplosiphon PCC7601) including repA for replication 
                     initiation of the plasmid in Fremyella diplosiphon PCC7601 
                     The device also carries the lacZalpha fragment for 
                     selection of assemblies carrying the device"
                     /db_xref="taxon:1531915"
     misc_feature    complement(146..404)
                     /note="lacZa-part2; lacZalpha fragment upstream of EcoICRI"
     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..2942
                     /note="CVD-CYA; device for the GC-adaptor assembly: 
                     cyanobacterial replicons"
     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"
     CDS             complement(585..1892)
                     /codon_start=1
                     /product="repA-pFDA"
                     /label=repA-pFDA
                     /note="repA-pFDA; derived from pPL2; 7; Fremyella 
                     diplosiphon PCC7601"
                     /protein_id="AII99656.1"
                     /translation="MKHTTFPHSNIIAQSTATEGENFVLQTRLHASILLLTPASKLWYL
                     CRALDLSGRGYVVLSPQNLELLQEKKSTIYRWLKDGKDLGLFRCYSWAGNTLKISLGSL
                     WRACKKARIKNWGTVANNIPLEEILKNNGRRTVATAMTIQDWQERSRYAAKNQLNPLER
                     KCFEIPATADVLTPQTSPKLTSGGTTGVLHVGEKRIFVGRSFIPFGVSQKRISNELNSQ
                     PASCGVSVRTVQRHVERLEVKHRQLMQARREYREIAHRIKQGATSWQCKSDADISFTWG
                     NQPDEIVLHERNGKSSARREGGHTIKLDQLCNYSNTHWRYRCNLYLLSYELTSMRTTRY
                     QWKKRGLNVKIAPVENCPSEISPQTPEMLESPKLHAEGRGLGGQNKSVKNENPQEVDTV
                     SNAFAPGGSEWAKTKAMLLAKQAERRQKRWDSLNSI"
     misc_feature    1152..1157
                     /label=MfeI
                     /note="MfeI"
     regulatory      complement(1893..2554)
                     /regulatory_class="promoter"
                     /note="PrepA; 662 nts upstream of repA-pFDA; derived from 
                     pPL2.7; Fremyella diplosiphon PCC7601"
     misc_feature    2417..2422
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2555..2560
                     /label=SacI
                     /note="SacI"
     regulatory      2561..2683
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     protein_bind    2574..2595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2610..2640
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      2646..2666
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    2648..2664
                     /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)."
     CDS             2685..2915
                     /codon_start=1
                     /product="lacZa"
                     /label=lacZa
                     /note="alpha fragment of lacZ GenBank: J01636 derived from 
                     Escherichia coli for alpha complementation in Escherichia 
                     coli blue/white screen; uninterrupted by a MCS"
                     /protein_id="AII99657.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLE"
     primer_bind     complement(2705..2721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2916..2921
                     /label=SacI
                     /note="SacI"
     misc_feature    2922..2942
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    2940..2945
                     /label=ZraI
                     /note="ZraI"
     misc_feature    complement(2946..3010)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    2964..2969
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(3006..3022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3011..3133)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(3028..3048)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    3030..3046
                     /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(3054..3084)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3099..3120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3349..4022)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(3408..3996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4167..5027)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99655.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5028..5132)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5028..5097)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    5158..5163
                     /label=ZraI
                     /note="ZraI"