LOCUS       Exported                3547 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD058, complete sequence.
ACCESSION   KM017885
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3547)
  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 3547)
  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 3547)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3547
                     /organism="Broad host range vector vector pCVD058"
                     /mol_type="other DNA"
                     /note="AhdI-yemGFP is a reporter device that comprises 
                     yemGFP downstream the AhdI restriction site"
                     /db_xref="taxon:1531916"
     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..1262)
                     /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="AII99659.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    671..676
                     /label=MfeI
                     /note="MfeI"
     misc_feature    1017..1022
                     /label=XbaI
                     /note="XbaI"
     misc_feature    complement(1242..1262)
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    1260..1265
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(1266..1330)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    1284..1289
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1326..1342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1331..1453)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(1348..1368)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    1350..1366
                     /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(1374..1404)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1419..1440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1669..2342)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(1728..2316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2487..3347)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99658.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3348..3452)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3348..3417)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    3478..3483
                     /label=ZraI
                     /note="ZraI"