pHRGFPTC vector (V005499)

Basic Vector Information

Vector Name:
pHRGFPTC
Antibiotic Resistance:
Ampicillin
Length:
9233 bp
Type:
Cloning vector
Replication origin:
pBBR1 oriV
Source/Author:
Quandt J, Hynes MF.
Promoter:
Pc

pHRGFPTC vector Map

pHRGFPTC9233 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200pBBR1 oriVpBBR1 RepSK primerKS primerSV40 NLSsmall t intronCmRyeGFPfrom pJQ200KSPc promoterCAP binding sitelac promoterlac operatorM13 revT3 promotertet promoterTcRAmpR

pHRGFPTC vector Sequence

LOCUS       40924_24962        9233 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHRGFPTC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9233)
  AUTHORS   Quandt J, Hynes MF.
  TITLE     Versatile suicide vectors which allow direct selection for gene 
            replacement in gram-negative bacteria
  JOURNAL   Gene 127 (1), 15-21 (1993)
  PUBMED    8486283
REFERENCE   2  (bases 1 to 9233)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Plasposons: modular self-cloning minitransposon derivatives for 
            rapid genetic analysis of gram-negative bacterial genomes
  JOURNAL   Appl. Environ. Microbiol. 64 (7), 2710-2715 (1998)
  PUBMED    9647854
REFERENCE   3  (bases 1 to 9233)
  AUTHORS   Ouahrani-Bettache S, Porte F, Teyssier J, Liautard JP, Kohler S.
  TITLE     pBBR1-GFP: a broad-host-range vector for prokaryotic promoter 
            studies
  JOURNAL   BioTechniques 26 (4), 620-622 (1999)
  PUBMED    10343896
REFERENCE   4  (bases 1 to 9233)
  AUTHORS   Ramos HJ, Roncato-Maccari LD, Souza EM, Soares-Ramos JR, Hungria M, 
            Pedrosa FO.
  TITLE     Monitoring Azospirillum-wheat interactions using the gfp and gusA 
            genes constitutively expressed from a new broad-host range vector
  JOURNAL   J. Biotechnol. 97 (3), 243-252 (2002)
  PUBMED    12084480
REFERENCE   5  (bases 1 to 9233)
  AUTHORS   Ramos HJO., Soares-Ramos JRL., Souza EM, Pedrosa FO.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2003) Department of Biochemistry and Molecular 
            Biology, Universidade Federal do Parana - UFPR, Centro Politecnico, 
            Curitiba, PR 81531-990, Brazil
REFERENCE   6  (bases 1 to 9233)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 9233)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1993"; volume: "127"; issue: "1"; pages: "15-21"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "1998"; volume: "64"; issue: "7"; pages:
            "2710-2715"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1999"; volume: "26"; issue: "4"; pages: 
            "620-622"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "J. 
            Biotechnol."; date: "2002"; volume: "97"; issue: "3"; pages: 
            "243-252"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (14-FEB-2003) Department of Biochemistry and Molecular Biology, 
            Universidade Federal do Parana - UFPR, Centro Politecnico, Curitiba,
            PR 81531-990, Brazil"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     2614..2630
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2664..2680)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3110..3130)
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     intron          complement(3260..3325)
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3494..4150)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(4216..4929)
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    4966..5465
                     /label=from pJQ200KS
                     /note="from pJQ200KS"
     promoter        complement(5473..5501)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    6010..6031
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6046..6076
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6084..6100
                     /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)."
     primer_bind     6108..6124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6145..6163
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6192..6221
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             6269..7456
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(8165..9022)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"

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