LOCUS       Exported               12351 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFTarget3-Tn7, complete sequence.
ACCESSION   JX644995
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12351)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     An improved method for the oriT-directed cloning and 
            functionalization of large bacterial genomic regions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12351)
  AUTHORS   Kvitko BH, McMillan IA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-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 12351)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12351
                     /organism="Cloning vector pFTarget3-Tn7"
                     /mol_type="other DNA"
                     /db_xref="taxon:1349685"
     CDS             1..375
                     /codon_start=1
                     /gene="ble"
                     /product="zeocin resistance protein"
                     /label=ble
                     /note="Ble"
                     /protein_id="AGO61969.1"
                     /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            1911..2448
                     /label=RK2/RP4 oriT
                     /note="RK2/RP4 oriT"
     oriT            2309..2417
                     /note="incP origin of transfer"
     CDS             complement(2624..3418)
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /EC_number="2.7.1.95"
                     /label=nptII
                     /note="NptII; kanamycin resistance"
                     /protein_id="AGO61970.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2624..3418)
                     /gene="nptII"
                     /label=nptII
     CDS             complement(3506..5317)
                     /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="AGO61971.1"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVAHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            complement(3506..5317)
                     /gene="uidA"
                     /label=uidA
     CDS             5675..6022
                     /codon_start=1
                     /gene="redF"
                     /product="RedF"
                     /label=redF
                     /protein_id="AGO61972.1"
                     /translation="MERRNRRTGRTEKARIWEVTDRTVRTWIGEAVAAAAADGVTFSVP
                     VTPHTFRHSYAMHMLYAGIPLKVLQSLMGHKSISSTEVYTKVFALDVAARHRVQFAMPE
                     SDAVAMLKQLS"
     gene            5675..6022
                     /gene="redF"
                     /label=redF
     rep_origin      6417..7031
                     /label=RK2/RP4 oriV
                     /note="RK2/RP4 oriV"
     rep_origin      7107..7326
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      7107..7323
                     /label=F ori2
                     /note="F ori2"
     CDS             7417..8172
                     /codon_start=1
                     /gene="repE"
                     /product="F plasmid replication initiation protein"
                     /label=repE
                     /note="RepE"
                     /protein_id="AGO61973.1"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     gene            7417..8172
                     /gene="repE"
                     /label=repE
     misc_feature    8175..8425
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             8751..9926
                     /codon_start=1
                     /gene="sopA"
                     /product="F plasmid partitioning protein A"
                     /label=sopA
                     /note="SopA"
                     /protein_id="AGO61974.1"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     gene            8751..9926
                     /gene="sopA"
                     /label=sopA
     CDS             9926..10897
                     /codon_start=1
                     /gene="sopB"
                     /product="F plasmid partitioning protein B"
                     /label=sopB
                     /note="SopB"
                     /protein_id="AGO61975.1"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     gene            9926..10897
                     /gene="sopB"
                     /label=sopB
     repeat_region   10970..11487
                     /label=sopC repeat region
                     /note="sopC repeat region"
     misc_feature    10970..11443
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    11698..11715
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     repeat_region   11716..11882
                     /label=Tn7L
                     /note="Tn7L"
     mobile_element  11716..11881
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     11893..11917
                     /label=Gateway attB1
                     /note="Gateway attB1"
     protein_bind    11893..11917
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    11918..11922
                     /label=target insert site
                     /note="target insert site"
     misc_recomb     11923..11947
                     /label=Gateway attB2
                     /note="Gateway attB2"
     protein_bind    complement(11923..11947)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     12069..12116
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(12069..12116)
                     /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)."
     misc_feature    12204..12283
                     /label=Mu strong gyrase site
                     /note="Mu strong gyrase site"
     promoter        12286..12333
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "