Basic Vector Information
- Vector Name:
- pFTCAPL-GW
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 11445 bp
- Type:
- Cloning vector
- Replication origin:
- oriV
- Source/Author:
- Kvitko BH, McMillan IA, Schweizer HP.
pFTCAPL-GW vector Vector Map
pFTCAPL-GW vector Sequence
LOCUS 40924_20606 11445 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pFTCAPL-GW, complete sequence.
ACCESSION .
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)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11445)
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
(04-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..11445
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 165..1337
/codon_start=1
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
CDS 1340..2308
/codon_start=1
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
misc_feature 2384..2857
/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
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
protein_bind 3307..3431
/label=attR1
/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..4196
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 4541..4843
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(4887..5011)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
oriT 5168..5276
/label=oriT
/note="incP origin of transfer"
CDS complement(5486..6277)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS complement(6368..8176)
/codon_start=1
/label=GUS
/note="beta-glucuronidase"
/translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVAHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
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=oriV
/note="origin of replication for the bacterial F plasmid"
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..11028
/codon_start=1
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
misc_feature 11034..11284
/label=incC
/note="incompatibility region of the bacterial F plasmid"
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