LOCUS       Exported                4989 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD050, complete sequence.
ACCESSION   KM017889
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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 4989)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4989
                     /organism="Broad host range vector vector pCVD050"
                     /mol_type="other DNA"
                     /note="pDC1Z is a replication device carrying a minimal 
                     fragment of pDC1 (an endogenous plasmid of Nostoc sp 
                     PCC8009) including repA for replication of the plasmid in 
                     Nostoc punctiforme The device also carries the lacZalpha 
                     fragment for selection of assemblies carrying the device"
                     /db_xref="taxon:1531908"
     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=EcoRV
                     /note="EcoRV"
     misc_feature    408..2704
                     /note="CVD-CYA; device for the GC-adaptor assembly: 
                     cyanobacterial replicons"
     misc_feature    408..428
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    429..434
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(600..1634)
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDC1; derived from pSUN202; Nostoc sp. PCC8009"
                     /protein_id="AII99670.1"
                     /translation="MANQNRAATQVEGLNHCPLTSQEDRSALSSNVPLPLVLILVKAEI
                     ILQPFMVRLHSGISRQPWARLWYLLRGFDDGSGRVKEIPLVLVEEILGVSQATVYRWLD
                     QGKAVGAFLSGYKVRIGLLTVYLGGLTKVAWHLNLKDWGVVAECPLWEVNANIRALTTG
                     IVTQRLQQRSRYAANRKLKPEYRKSYGAPHPNELLGDEGQSSFKLGAGEVPFVLHVSPS
                     RVFVSKSFIHYGTSQNAVSCKLGIHTRTVRRHQRTLGMEKRQLCQAKYEYAQLDRAVSN
                     EASECWAWSSEGTQSDIGYQSLGISHWVKGRSRFLMASCQEPESSSRTLTSCQRANCLG
                     VSSK"
     misc_feature    1221..1228
                     /label=SwaI
                     /note="SwaI"
     regulatory      complement(1635..2316)
                     /regulatory_class="promoter"
                     /note="PrepA; 682 nts upstream of repA-pDC1; derived from 
                     pSUN202; Nostoc sp. PCC8009"
     misc_feature    2113..2118
                     /label=MfeI
                     /note="MfeI"
     misc_feature    2317..2322
                     /label=SacI
                     /note="SacI"
     regulatory      2323..2445
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     protein_bind    2336..2357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2372..2402
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      2408..2428
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    2410..2426
                     /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             2447..2677
                     /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="AII99671.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLE"
     primer_bind     complement(2467..2483)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2678..2683
                     /label=SacI
                     /note="SacI"
     misc_feature    2684..2704
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons or Escherichia coli origins to be 
                     assembled with a cyanobacterial replicon; synthetic"
     misc_feature    2702..2707
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(2708..2772)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    2726..2731
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2768..2784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2773..2895)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(2790..2810)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    2792..2808
                     /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(2816..2846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2861..2882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3111..3784)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3929..4789)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99669.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4790..4894)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(4790..4859)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    4920..4925
                     /label=ZraI
                     /note="ZraI"