LOCUS       Exported                7744 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pXF20pemIK-GW, complete sequence.
ACCESSION   KX036765
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  AUTHORS   Burbank L, Stenger DC.
  TITLE     Plasmid vectors for Xylella fastidiosa utilizing a toxin-antitoxin 
            system for plasmid stability in the absence of antibiotic selection
  JOURNAL   Phytopathology (2016) In press
  PUBMED    27088393
REFERENCE   2  (bases 1 to 7744)
  AUTHORS   Burbank LP, Stenger DC.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2016) Crop Diseases Pests and Genetics Research 
            Unit, USDA Agricultural Research Service, 9611 S Riverbend Ave, 
            Parlier, CA 93648, USA
REFERENCE   3  (bases 1 to 7744)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7744
                     /organism="Cloning vector pXF20pemIK-GW"
                     /mol_type="other DNA"
                     /db_xref="taxon:1840504"
     rep_origin      1..619
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    893..914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        929..959
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    967..983
                     /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     991..1007
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1104..1488
     CDS             complement(1527..2393)
                     /codon_start=1
                     /gene="trfA"
                     /product="replication initiator protein"
                     /label=trfA
                     /protein_id="ANA75341.1"
                     /translation="MSSIELSLTKMRARQEANLQKSLLQQLQLPIWTDAQRGVPNSLLR
                     SALFAAIQRGKRPYLKRQVMASVENASITYTGIRLCQDYLTVWECLVHVAREQQLGNRC
                     EVTTYQLLRLLDKKDTGGNRRVLRRQLAELQATALEVRQGRYTYSGSLIDEAYHDEETG
                     RAIIVLNQRIVALYQSDGYTRVSWQIRCDLNTSLAKWLHGFYATHRQPYPYSVAKIHEL
                     CGSAAKSLKDFQEKTLAPALEELASVCKNHGEHFSYMIERRPGGKSIVYVTRDVGTKLP
                     LTPEICG"
     gene            complement(1527..2393)
                     /gene="trfA"
                     /label=trfA
     CDS             2435..2446
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_recomb     2484..2608
                     /label=attR
                     /note="attR"
     protein_bind    2484..2608
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2633..2663
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2717..3376
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="ANA75340.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            2717..3376
                     /gene="cat"
                     /label=cat
     CDS             3718..4023
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB killer protein"
                     /label=ccdB
                     /protein_id="ANA75342.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3718..4023
                     /gene="ccdB"
                     /label=ccdB
     protein_bind    complement(4064..4188)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     4087..4164
                     /label=attR
                     /note="attR"
     CDS             complement(4259..4621)
                     /codon_start=1
                     /gene="pemK"
                     /product="mRNA interferase"
                     /label=pemK
                     /protein_id="ANA75343.1"
                     /translation="MGRCSSSRTRTDMKRGDVYMVDLEPTAGHEQRGHRPVVVISSERF
                     NRLTSCPVILPITNGGEFASRLGFAVELLGTITTGVVRCDQPRALDLLARNARKVESLP
                     PNILADVLAKAVTIFQ"
     gene            complement(4259..4621)
                     /gene="pemK"
                     /label=pemK
     CDS             complement(4582..4845)
                     /codon_start=1
                     /gene="pemI"
                     /product="antitoxin"
                     /label=pemI
                     /protein_id="ANA75344.1"
                     /translation="MHTTNLRKVGGSIMLAVPPAFLDQLHLEVGATVGLAVTDGRLVIE
                     PVLHPQYTLDQLLAEAEASGAYPLPPEEREWVDAPAVGRELI"
     gene            complement(4582..4845)
                     /gene="pemI"
                     /label=pemI
     promoter        complement(4964..4982)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4989..5005)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5146..5574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             5918..6712
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase II"
                     /label=nptII
                     /note="kanamycin/neomycin resistance"
                     /protein_id="ANA75345.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            5918..6712
                     /gene="nptII"
                     /label=nptII
     CDS             6730..7590
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="ANA75346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            6730..7590
                     /gene="bla"
                     /label=bla