LOCUS       Exported                3400 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD012, complete sequence.
ACCESSION   KM017936
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3400)
  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 3400)
  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 3400)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3400
                     /organism="Broad host range vector vector pCVD012"
                     /mol_type="other DNA"
                     /note="nptII_A7120 is a device that confers kanamycin or 
                     neomycin (Km/Nm) resistance"
                     /db_xref="taxon:1531870"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    370..375
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(373..1425)
                     /note="CVD-ANTR; device for the GC-adaptor assembly: 
                     antibiotic resistance markers"
     misc_feature    complement(373..393)
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    394..399
                     /label=AgeI
                     /note="AgeI"
     regulatory      406..562
                     /regulatory_class="promoter"
                     /note="PnptII; upstream region of nptII ORF including 
                     promoter and RBS in INSD accession V00618; derived from 
                     pAM505"
     CDS             563..1354
                     /codon_start=1
                     /product="nptII_A7120"
                     /label=nptII_A7120
                     /note="confers kanamycin resistance; aminoglycoside 
                     phosphotransferase"
                     /protein_id="AII99780.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      1363..1389
                     /regulatory_class="terminator"
                     /note="trpA T; trpA terminator from Lorist6 INSD accession 
                     X98450, one nt shorter than reference; Lorist6"
     misc_feature    1399..1404
                     /label=NheI
                     /note="NheI"
     misc_feature    complement(1405..1425)
                     /note="GC; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers or Escherichia coli origins 
                     to be assembled with a cyanobacterial replicon; synthetic"
     misc_feature    1423..1428
                     /label=EcoRV
                     /note="EcoRV"
     rep_origin      complement(1594..2267)
                     /direction=LEFT
                     /note="colE1_pMA-RQ; origin of replication of colE1 found 
                     inderived from pMA-RQ GeneART vector"
     rep_origin      complement(1653..2241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2412..3272)
                     /codon_start=1
                     /product="bla_pMA-RQ"
                     /label=bla_pMA-RQ
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pMA-RQ GeneART vector, has one point mutation 
                     compared to pUC19"
                     /protein_id="AII99779.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3273..3377)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3273..3342)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"