LOCUS       sequence-245919-        6033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for gene expression in Xylella fastidiosa, includes pemI/pemK
            toxin-antitoxin system for plasmid stability.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6033)
  AUTHORS   Burbank LP, Stenger DC
  TITLE     Plasmid Vectors for Xylella fastidiosa Utilizing a Toxin-Antitoxin 
            System for Stability in the Absence of Antibiotic Selection.
  JOURNAL   Phytopathology. 2016 Aug;106(8):928-36. doi: 
            10.1094/PHYTO-02-16-0097-R. Epub 2016 Jun 7.
  PUBMED    27088393
REFERENCE   2  (bases 1 to 6033)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1094/PHYTO-02-16-0097-R"; journalName: "Phytopathology"; date: 
            "2016-08"; volume: "106"; issue: "8"; pages: "928-36"
COMMENT     Sequence Label: pXf20pemIK
FEATURES             Location/Qualifiers
     source          1..6033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     443..462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     696..713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    830..851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(866..871,872..889,890..896)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    904..920
                     /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     909..931
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     928..944
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     928..944
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2372..2383
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3189..3208)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3190..3208)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3215..3232)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3215..3231)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3246)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3372..3800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3459..3478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3669..3690
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             4144..4938
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4198..4217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4808..4827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4956..5024,5025..5816)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5174..5193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5987..6033,1..542)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
       61 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      661 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      721 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      781 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      841 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      901 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      961 ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt cggcttccat
     1021 aaagttacat gaaacggatt ttttataaaa aatggtttgt gaaagcgcag tgtgggcaaa
     1081 agcacgctta agcgtgctgg tgcacgtagt gcattgtcca tacggagcgt taagcggtgg
     1141 agcttgtggg cagagacttg cggcggtggg caaccgcttg cggttgtcca cggacgtgag
     1201 tcggtgtgcg tagcgcactg tccacaaatc cacggcttta gttaaatccg ttgctttaga
     1261 ttagaaaaag attaaagaag tattagaggt tatccacagg caacgtaaga tattgatttt
     1321 atagttaaat tttcttattt ttcgtgaaaa ttttaggatg ctgcgacggt aaaaaggcga
     1381 atgctgcgac ggtaaaatct aggattctgc gacggtacat gaggatgctg cgacggtaaa
     1441 aaggcgagtg ctgcgacggt aagttatcca cagatttcag gggttaatgg cagcttggtt
     1501 cctacgtcac gggtaacgta tacaattgac ttaccacctg ggcggcgctc aatcatgtag
     1561 ctgaagtgct ctccgtggtt cttacaaacg cttgccagct cttcaagtgc tggagctaac
     1621 gtcttttctt ggaagtcctt cagagactta gctgcgctgc cgcacagctc atgaatcttg
     1681 gccacactgt agggataggg ttgacggtga gttgcataga atccatgaag ccacttggcc
     1741 agagatgtat ttagatcaca tctaatctgc caagaaaccc tcgtgtagcc gtcgctttgg
     1801 tacaaggcaa cgatgcgttg atttagtaca atgatggcgc ggcctgtttc ctcatcatga
     1861 taggcttcgt cgatcaagct gccagagtac gtgtagcgtc cttgtcgtac ttcaagcgcg
     1921 gttgcttgca gctcggcaag ttgtcgtcgt agcactcgcc gattgccacc tgtgtccttc
     1981 ttgtccaaga gtctcaagag ttgataggta gtgacttcgc agcgattgcc taactgctgt
     2041 tcgcgtgcga cgtgaacaag gcattcccac acagtcaaat agtcctgaca taagcggatt
     2101 ccggtgtacg tgatggatgc gttttcaacg ctggccatca cttggcgctt gaggtagggg
     2161 cgtttaccgc gttgaatcgc ggcaaagagc gccgatcgaa gtaagctgtt cggaacgccg
     2221 cgctgcgcat ctgtccaaat tggcaattga agctgctgca ataggctctt ttgcaggttg
     2281 gcttcctgtc tggcacgcat tttcgttagt gaaagttcta tgctgctcac gcttcctcct
     2341 tggttgtgta gtcacgctca agaacaactc gatagaaggt cgagcgcgtg atgttgtata
     2401 agccgaattc tgcagatatc catcacactg gcggccgcga attcactagt gattggttgc
     2461 agtggcctga acttgcccca aaaattactg gaagatggtg acagcttttg ccaagacatc
     2521 ggccaagatg tttggtggta ggctttccac tttgcgggca ttgcgagcga gtaagtcgag
     2581 cgcacgtggt tgatcacatc gaacgacgcc cgttgtaatc gtacctagta attccacagc
     2641 gaacccgagg cgggaggcga actcgccgcc gtttgtgatt ggaaggatca ccggacaaga
     2701 ggtaagccga ttgaatcgct cactggagat aacaaccacc ggacggtgac cgcgttgttc
     2761 gtgtccggcg gttggctcca gatcgaccat gtaaacatcg ccgcgtttca tatcagttcg
     2821 cgtcctactg ctggagcatc gacccattca cgttcctcgg gcggcaacgg gtaagccccg
     2881 gaggcttccg cttcggccag cagttgatcc agcgtgtact gaggatgtag gactggctca
     2941 atgaccagac gaccgtctgt gacggctaag ccgaccgttg cgccgacctc aagatgaagc
     3001 tgatcaagga aagcgggggg aacggccagc atgatcgatc caccgacttt acgaaggttt
     3061 gtggtgtgca tgatgggctt ctttcgttaa gttatattaa agtataatat aattcctcca
     3121 tttatacgct tatagcgaaa atcgaattcc cgcggccgct cgagcatgca tctagagggc
     3181 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     3241 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3301 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3361 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3661 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3721 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3781 acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca agggctgcta aaggaagcgg
     3841 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     3901 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     3961 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     4021 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     4081 ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt
     4141 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4201 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4261 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4321 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4381 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4441 caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4501 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4561 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4621 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4681 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4741 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4801 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4861 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4921 cttgacgagt tcttctgaat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4981 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5041 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5101 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5161 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5221 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5281 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5341 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5401 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5461 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5521 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5581 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5641 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5701 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5761 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5821 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5881 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6001 ttctgcgcgt aatctgctgc ttgcaaacaa aaa
//
