LOCUS       Exported                4866 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD056, complete sequence.
ACCESSION   KM017883
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  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 4866)
  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 4866)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4866
                     /organism="Broad host range vector vector pCVD056"
                     /mol_type="other DNA"
                     /note="pFDA 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"
                     /db_xref="taxon:1531914"
     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..2581
                     /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="AII99654.1"
                     /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=MfeI
                     /note="MfeI"
     misc_feature    2561..2581
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    2579..2584
                     /label=ZraI
                     /note="ZraI"
     misc_feature    complement(2585..2649)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    2603..2608
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2645..2661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2650..2772)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(2667..2687)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    2669..2685
                     /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(2693..2723)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2738..2759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2988..3661)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(3047..3635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3806..4666)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99653.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4667..4771)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(4667..4736)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    4797..4802
                     /label=ZraI
                     /note="ZraI"