pUCTCAPR3-GW vector (V002351)

LOCUS       40924_45403        8522 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCTCAPR3-GW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8522)
  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 8522)
  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 8522)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8522)
  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..8522
                     /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(8240..8287)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    8375..8454
                     /label=Mu strong gyrase site
                     /note="Mu strong gyrase site"
     promoter        8457..8504
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"