pCVD052 vector (V001535)

Basic Vector Information

Vector Name:
pCVD052
Antibiotic Resistance:
Ampicillin
Length:
6852 bp
Type:
Broad host range vector
Replication origin:
ori
Source/Author:
Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.

pCVD052 vector Map

pCVD0526852 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600M13 fwdC3G3; GC-adaptor for seamless assembly, flanking cyanobacterial replicons; syntheticSacIlacZ-alphaPlacZa; lacI-lacZ intergenic region derived from Escherichia coli including the promoter and RBSSacIPrepA; PrepA; upstream region of repA of pDU1 including promoter and ribosomal binding site,derived from pAM505; Nostoc sp. PCC7524repArho-ITTS; rho-ITTS; rho-independent transcriptional termination signal, downstream of repA in pDU1, from pAM505; Nostoc sp. PCC7524XbaIorfCXbaIhyp1XbaIXbaIG5C5; GC-adaptor for seamless assembly, flanking a BHR replicons, neutral sites, or cyanobacterial replicons; syntheticXbaIM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterZraI

pCVD052 vector Sequence

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"

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