LOCUS       Exported                4989 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD059, complete sequence.
ACCESSION   KM017886
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 pCVD059"
                     /mol_type="other DNA"
                     /note="catccdB_AhdI-yemGFP is a reporter device where 
                     yemGFP is downstream of the ccdB gene and a chloramphenicol
                     resistance marker, flanked by AhdI restriction sites"
                     /db_xref="taxon:1531917"
     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    complement(408..2704)
                     /note="CVD-cassette; device for the GC-adaptor assembly: 
                     expression/reporter cassette, promoter-reporter module, 
                     custom sequence"
     misc_feature    complement(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"
     regulatory      complement(441..469)
                     /regulatory_class="terminator"
                     /note="rrnB T2; rrnB T2; rrnB T2 terminator partial 
                     sequence"
     terminator      442..469
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    478..502
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(519..1235)
                     /codon_start=1
                     /product="yemGFP-gc"
                     /label=yemGFP-gc
                     /note="expression plasmid containing yemGFP, a GFP allele 
                     with F64L, S65T, and A206K mutations and that has been 
                     codon-optimized for expression in yeast; the first two 
                     mutations correspond to GFPmut1, and the third mutation 
                     interferes with GFP dimerization; optimized codons at the 
                     5-end 18nts to increase GC"
                     /protein_id="AII99663.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    671..676
                     /label=MfeI
                     /note="MfeI"
     misc_feature    1017..1022
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(1282..1587)
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /note="from a destination cassette used in the Gateway 
                     Technology developed by Life Technologies"
                     /protein_id="AII99662.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      complement(1843..1892)
                     /regulatory_class="promoter"
                     /note="PccdB; upstream region of the ccdB ORF including 
                     promoter and RBS; derived from the destination cassette 
                     used in the Gateway Technology developed by Life 
                     Technologies"
     CDS             complement(1929..2588)
                     /codon_start=1
                     /product="cat"
                     /label=cat
                     /note="confers chloramphenicol resistance; chloramphenicol 
                     acetyltransferase; derived from a destination cassette used
                     in the Gateway Technology developed by Life Technologies"
                     /protein_id="AII99660.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      complement(2589..2672)
                     /regulatory_class="promoter"
                     /note="Pcat; upstream region of the cat ORF including 
                     promoter and RBS; derived from the destination cassette 
                     used in the Gateway Technology developed by Life 
                     Technologies; Life Technologies"
     promoter        complement(2642..2672)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    complement(2684..2704)
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; 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="AII99661.1"
                     /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"