LOCUS       Exported                8382 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCTCAPR2-GW, complete sequence.
ACCESSION   JX573199
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8382
                     /organism="Cloning vector pUCTCAPR2-GW"
                     /mol_type="other DNA"
                     /label=oriTn7 right capture vector
                     /note="oriTn7 right capture vector"
                     /db_xref="taxon:1332829"
     CDS             1..375
                     /codon_start=1
                     /gene="ble"
                     /product="zeocin resistance protein"
                     /label=ble
                     /note="Ble"
                     /protein_id="AGM33182.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            1..375
                     /gene="ble"
                     /label=ble
     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      586..672
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      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
                     /gene="Escherichia coli rrnB"
                     /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            1917..2764
                     /label=RK2/RP4 oriT
                     /note="RK2/RP4 oriT"
     oriT            2309..2418
                     /note="incP origin of transfer"
     terminator      3114..3200
                     /gene="Escherichia coli rrnB"
                     /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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3892..4425)
                     /codon_start=1
                     /gene="aacCI"
                     /product="gentamicin 3-N-acetyltransferase"
                     /EC_number="2.3.1.60"
                     /label=aacCI
                     /note="AacCI; gentamicin resistance protein"
                     /protein_id="AGM33183.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(3892..4425)
                     /gene="aacCI"
                     /label=aacCI
     promoter        complement(4614..4642)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5362..6035
                     /label=pUC replication origin
                     /note="pUC replication origin"
     rep_origin      5365..5953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6241..6262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     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"
     misc_recomb     6414..6538
                     /label=Gateway attR1
                     /note="Gateway attR1"
     protein_bind    6414..6538
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /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..7306
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /EC_number="2.3.1.28"
                     /label=cat
                     /note="Cat; chloramphenicol resistance protein"
                     /protein_id="AGM33184.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            6647..7306
                     /gene="cat"
                     /label=cat
     CDS             7648..7953
                     /codon_start=1
                     /gene="ccdB"
                     /product="GyrA toxin"
                     /label=ccdB
                     /note="CcdB; Gateway counter-selection toxin"
                     /protein_id="AGM33185.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            7648..7953
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     7994..8118
                     /label=Gateway attR2
                     /note="Gateway attR2"
     protein_bind    complement(7994..8118)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     8180..8227
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(8180..8227)
                     /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)."
     promoter        8317..8364
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "