LOCUS       Exported                4632 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD016, complete sequence.
ACCESSION   KM017928
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4632)
  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 4632)
  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 4632)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4632
                     /organism="Broad host range vector vector pCVD016"
                     /mol_type="other DNA"
                     /note="cPtrc-gateway-dest is an expression device 
                     comprising a trc promoter, but not the lacIq gene, upstream
                     of a Gateway destination cassette"
                     /db_xref="taxon:1531874"
     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..2347
                     /note="CVD-cassette; device for the GC-adaptor assembly: 
                     expression/reporter cassette, promoter-reporter module, 
                     custom sequence"
     misc_feature    408..428
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    429..434
                     /label=AgeI
                     /note="AgeI"
     regulatory      454..482
                     /regulatory_class="terminator"
                     /note="lpp T; lpp T; lpp terminator"
     regulatory      491..564
                     /regulatory_class="promoter"
                     /note="Ptrc; trc promoter, ends defined by the overlap 
                     between pTrcHis2-A for the 5 end and pAM2255 for the 3 
                     end; synthetic"
     promoter        494..523
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      529..549
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    531..547
                     /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)."
     misc_recomb     572..696
                     /note="attR1; recombination site for the LR reaction used 
                     in the Gateway Technology developed by Life Technologies; 
                     Life Technologies; lambda"
     protein_bind    572..696
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      721..804
                     /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        721..751
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             805..1464
                     /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="AII99762.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      1501..1550
                     /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             1806..2111
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /note="from a destination cassette used in the Gateway 
                     Technology developed by Life Technologies"
                     /protein_id="AII99764.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     2152..2276
                     /note="attR2; recombination site for the LR reaction used 
                     in the Gateway Technology developed by Life Technologies; 
                     Life Technologies; lambda"
     protein_bind    complement(2152..2276)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2285..2313
                     /regulatory_class="terminator"
                     /note="rrnB T2; rrnB T2; rrnB T2 terminator partial 
                     sequence"
     terminator      2285..2312
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    2321..2326
                     /label=XbaI
                     /note="XbaI"
     misc_feature    2327..2347
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    2345..2350
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(2351..2415)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    2369..2374
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2411..2427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2416..2538)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(2433..2453)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    2435..2451
                     /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(2459..2489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2504..2525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2754..3427)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3572..4432)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99763.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4433..4537)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(4433..4502)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    4563..4568
                     /label=ZraI
                     /note="ZraI"