pUCTCAPR2-GW vector (V002352)

Basic Vector Information

      • Vector Name:
      • pUCTCAPR2-GW
      • Antibiotic Resistance:
      • Chloramphenicol
      • Length:
      • 8382 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Kvitko BH, McMillan IA, Schweizer HP.

pUCTCAPR2-GW vector Vector Map

pUCTCAPR2-GW8382 bp400800120016002000240028003200360040004400480052005600600064006800720076008000BleoRFRTrrnB T1 terminatorlambda t0 terminatorKS primerTn7RI-SceI restriction siteM13 fwdrrnB T1 terminatorrrnB T2 terminatororiTrrnB T1 terminatorrrnB T2 terminatorAmpR promoterGmRPc promoteroriCAP binding sitelac promoterlac operatorM13 revI-SceI restriction siteattR1lac UV5 promoterCmRccdBattR2FRTEM7 promoter

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pUCTCAPR2-GW vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_45398        8382 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCTCAPR2-GW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8382)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     An Improved Method for oriT-Directed Cloning and Functionalization 
            of Large Bacterial Genomic Regions
  JOURNAL   Appl. Environ. Microbiol. 79 (16), 4869-4878 (2013)
  PUBMED    23747708
REFERENCE   2  (bases 1 to 8382)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-SEP-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   3  (bases 1 to 8382)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8382)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2013"; volume: "79"; issue: "16"; 
            pages: "4869-4878"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-SEP-2012) Microbiology Immunology and Pathology, Colorado State 
            University, IDRC at Foothills Campus, Campus Delivery 0922, Fort 
            Collins, CO 80523-0922, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..372
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_recomb     486..533
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(486..533)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(586..672)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(775..869)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(897..913)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   918..1116
                     /label=Tn7R
                     /note="Tn7R"
     misc_feature    1264..1281
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     primer_bind     complement(1291..1307)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1657..1743
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1835..1862
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     oriT            2309..2418
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(3117..3160)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3292..3319
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3338..3429
                     /label=AmpR promoter
     CDS             complement(3895..4425)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(4614..4642)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5365..5953
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6241..6262
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6277..6307
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6315..6331
                     /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     6339..6355
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6374..6391
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     protein_bind    6414..6538
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6563..6593
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6647..7303
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             7648..7950
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(7994..8118)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(8180..8227)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        8317..8364
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"

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