LOCUS       Exported               11445 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFTCAPL-GW, complete sequence.
ACCESSION   JX573201
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11445)
  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 11445)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-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 11445)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11445
                     /organism="Cloning vector pFTCAPL-GW"
                     /mol_type="other DNA"
                     /label=oriTn7 left capture vector
                     /note="oriTn7 left capture vector"
                     /db_xref="taxon:1332828"
     CDS             165..1340
                     /codon_start=1
                     /gene="sopA"
                     /product="F plasmid partitioning protein A"
                     /label=sopA
                     /note="SopA"
                     /protein_id="AGM33190.1"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     gene            165..1340
                     /gene="sopA"
                     /label=sopA
     CDS             1340..2311
                     /codon_start=1
                     /gene="sopB"
                     /product="F plasmid partitioning protein B"
                     /label=sopB
                     /note="SopB"
                     /protein_id="AGM33191.1"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     gene            1340..2311
                     /gene="sopB"
                     /label=sopB
     repeat_region   2384..2901
                     /label=sopC repeat region
                     /note="sopC repeat region"
     misc_feature    2384..2857
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    3112..3129
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     mobile_element  3130..3295
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     3307..3431
                     /label=Gateway attR1
                     /note="Gateway attR1"
     protein_bind    3307..3431
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3456..3486
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3540..4199
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /EC_number="2.3.1.28"
                     /label=cat
                     /note="Cat; chloramphenicol resistance protein"
                     /protein_id="AGM33192.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            3540..4199
                     /gene="cat"
                     /label=cat
     CDS             4541..4846
                     /codon_start=1
                     /gene="ccdB"
                     /product="GyrA toxin"
                     /label=ccdB
                     /note="CcdB; Gateway counter-selection toxin"
                     /protein_id="AGM33193.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            4541..4846
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     4887..5011
                     /label=Gateway attR2
                     /note="Gateway attR2"
     protein_bind    complement(4887..5011)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     oriT            5029..5307
                     /label=RK2/RP4 oriT
                     /note="RK2/RP4 oriT"
     oriT            5168..5276
                     /note="incP origin of transfer"
     CDS             complement(5483..6277)
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /EC_number="2.7.1.95"
                     /label=nptII
                     /note="NptII; kanamycin resistance"
                     /protein_id="AGM33194.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(5483..6277)
                     /gene="nptII"
                     /label=nptII
     CDS             complement(6365..8176)
                     /codon_start=1
                     /gene="uidA"
                     /product="beta-glucuronidase"
                     /EC_number="3.2.1.31"
                     /label=uidA
                     /note="UidA; cleaves 5-bromo-4-chloro-3-indolyl glucuronide
                     (X-gluc) colorimetric marker"
                     /protein_id="AGM33195.1"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVAHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            complement(6365..8176)
                     /gene="uidA"
                     /label=uidA
     CDS             8534..8881
                     /codon_start=1
                     /gene="redF"
                     /product="RedF"
                     /label=redF
                     /protein_id="AGM33196.1"
                     /translation="MERRNRRTGRTEKARIWEVTDRTVRTWIGEAVAAAAADGVTFSVP
                     VTPHTFRHSYAMHMLYAGIPLKVLQSLMGHKSISSTEVYTKVFALDVAARHRVQFAMPE
                     SDAVAMLKQLS"
     gene            8534..8881
                     /gene="redF"
                     /label=redF
     rep_origin      9276..9890
                     /label=RK2/RP4 oriV
                     /note="RK2/RP4 oriV"
     rep_origin      9966..10185
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      9966..10182
                     /label=F ori2
                     /note="F ori2"
     CDS             10276..11031
                     /codon_start=1
                     /gene="repE"
                     /product="F plasmid replication initiation protein"
                     /label=repE
                     /note="RepE"
                     /protein_id="AGM33197.1"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     gene            10276..11031
                     /gene="repE"
                     /label=repE
     misc_feature    11034..11284
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"