LOCUS       sequence-189220-        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector for allelic exchange in Listeria utilizing an 
            engineered counterselection marker for Listeria monocytogenes based 
            on a mutated phenylalanyl-tRNA synthetase gene (pheS*)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  AUTHORS   Argov T, Rabinovich L, Sigal N, Herskovits AA
  TITLE     An Effective Counterselection System for Listeria monocytogenes and 
            Its Use To Characterize the Monocin Genomic Region of Strain 10403S.
  JOURNAL   Appl Environ Microbiol. 2017 Mar 2;83(6). pii: e02927-16. doi: 
            10.1128/AEM.02927-16. Print 2017 Mar 15.
  PUBMED    28039138
REFERENCE   2  (bases 1 to 7703)
  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"; journalName: "Appl 
            Environ Microbiol."; date: "2017-03-2"; volume: "83(6). pii: 
            e02927-16. doi"; pages: " 10.1128/AEM.02927-16. Print 2017 Mar 15"
COMMENT     Sequence Label: pLR16-Phes*
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..50)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     138..154
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     139..155
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(189..205)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(189..205)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     oriT            1613..1722
                     /note="incP origin of transfer"
     protein_bind    4257..4278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4293..4298,4299..4316,4317..4323)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4331..4347
                     /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     4336..4358
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4355..4371
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4355..4371
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(4560..5219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5134..5153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5220..5322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     5550..5572
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6994..7011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7128..7673)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(7208..7227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctagagcgg ccgccaccgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
       61 aatgtcatga taataatggt ttcttagacg tcattaaccc tcactaaagg gaacaaaagc
      121 tggtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca
      181 gcccggggga tccactagtt ctagagcggc cgccaccgat gtgaggcatt ttcgctcttt
      241 ccggcgaggc tagttaccct taagttattg gtatgactgg ttttaagcgc aaaaaaagtt
      301 gctttttcgt acctattaat gtatcgttag aaaaccgact gtaaaaagta cagtcggcat
      361 tatctcatat tataaaagcc agtcattagg cctatctgac aattcctgaa tagagttcat
      421 aaacaatcct gcatgataac catcacaaac agaatgatgt acctgtaaag atagcggtaa
      481 atatattgaa ttacctttat taatgaattt tcctgctgta ataatgggta gaaggtaatt
      541 actattatta ttgatattta agttaaaccc agtaaatgaa gtccatggaa taatagaaag
      601 agaaaaagca ttttcaggta taggtgtttt gggaaacaat ttccccgaac cattatattt
      661 ctctacatca gaaaggtata aatcataaaa ctctttgaag tcattcttta caggagtcca
      721 aataccagag aatgttttag atacaccatc aaaaattgta taaagtggct ctaacttatc
      781 ccaataacct aactctccgt cgctattgta accagttcta aaagctgtat ttgagtttat
      841 cacccttgtc actaagaaaa taaatgcagg gtaaaattta tatccttctt gttttatgtt
      901 tcggtataaa acactaatat caatttctgt ggttatacta aaagtcgttt gttggttcaa
      961 ataatgatta aatatctctt ttctcttcca attgtctaaa tcaattttat taaagttcat
     1021 ttgatatgcc tcctaaattt ttatctaaag tgaatttagg aggcttactt gtctgctttc
     1081 ttcattagaa tcaatccttt tttaaaagtc aatattactg taacataaat atatatttta
     1141 aaaatatccc actttatcca attttcgttt gttgaactaa tgggtgcttt agttgaagaa
     1201 taaaagacca cattaaaaaa tgtggtcttt tgtgtttttt taaaggattt gagcgtagcg
     1261 aaaaatcctt ttctttctta tcttgataat aagggtaact attgccggga tagactgtaa
     1321 cattctcacg cataaaatcc cctttcattt tctaatgtaa atctattacc ttattattaa
     1381 ttcaattcgc tcataattaa tcctttttct tattacgcaa aatggcccga tttaagcaca
     1441 ccctttattc cgttaatgcg ccatgacagc catgataatt actaatacta ggagaagtta
     1501 ataaatacgt atcagtgcaa tttattcttc aaatgtagca cctgaagtca gccccatacg
     1561 atataagttg taattctccg ccgcttgccc tcatctgtta cgccggcggt agccggccag
     1621 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag
     1681 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata
     1741 caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat
     1801 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc
     1861 tgcttccctg ctgttttgtg gaatatctac cgactggaaa caggcaaatg caggaaatta
     1921 ctgaactgag gggacaggcg agaggcgatc gcataacttc gtatagcata catgcataaa
     1981 ctgcatccct taacttgttt ttcgtgtgcc tattttttgt gaatcgatta tgtcttttgc
     2041 gcagtcggct taaaccagtt ttcgttggtg cgaaaaaaga gtgtcttgtg acactcttaa
     2101 attcaaaatc tatcggtcag atttataccg atttgatttt atatattctt gaataacata
     2161 cgccgagtta tcacataaaa gcgggaacca atcatcaaat ttaaacttca ttgcataatc
     2221 cattaaactc ttaaattcta cgattccttg ttcatcaata aactcaatca tttctttaat
     2281 taatttatat ctatctgttg ttgttttctt taataattca tcaacatcta caccgccata
     2341 aactatcata tcttcttttt gatatttaaa tttattagga tcgtccatgt gaagcatata
     2401 tctcacaaga cctttcacac ttcctgcaat ctgcggaata gtcgcattca attcttctgt
     2461 aattattttt atctgttcat aagatttatt accctcatac atcactagaa tatgataatg
     2521 ctcttttttc atcctatctt ctgtatcagt atccctatca tgtaatggag acactacaaa
     2581 ttgaatgtgt aactctttta aatactctaa ccactcggct tttgctgatt ctggatataa
     2641 aacaaatgtc caattacgtc ctcttgaatt tttcttgttt tcagtttctt ttattacatt
     2701 ttcgctcatg atataataac ggtgctaata catttaacaa aatttagtca tagataggca
     2761 gcatgccagt gctgtctatc tttttttgtt taaaatgcac cgtattcctc ctttgcatat
     2821 ttttttatta gaataccggt tgcatctgat ttgctaatat tatatttttc tttgattcta
     2881 tttaatatct cattttcttc tgttgtaagt cttaaagtaa cagcaacttt tttctcttct
     2941 tttctatcta caaccatcac tgtacctccc aacatctgtt tttttcactt taacataaaa
     3001 aacaaccttt taacattaaa aacccaatat ttatttattt gtttggacaa tggacaatgg
     3061 acacctaggg gggaggtcgt agtacccccc tatgttttct cccctaaata accccaaaaa
     3121 tctaagaaaa aaagacctca aaaaggtctt taattaacat ctcaaatttc gcatttattc
     3181 caatttcctt tttgcgtgtg atgcgctgcg tccattaaaa atcctagagc tttgcaaccg
     3241 aaagttaata gctgtcgcta ctactttcgc ttacgctcta agtatatttt aaggactgtc
     3301 acacgcaaaa agttttctcg gcataaaagt acctctacat ctctaaatcg tctgtacgct
     3361 gtttctcacg ctttctatcg accttctgga cattatcctg tacaacatcc ataaactgtc
     3421 ccacacgctc gaatttggaa tcattaaaga atttctcttt aagcctatta aaccctttct
     3481 caaacccagg gaaattcgcc ctcgcagcac gatataaagt cactgtacta tcttgaaatt
     3541 tctctgatac attcaactgc tcattcaaac tatcattctc tcgctttaat ttattaacct
     3601 ctttactttt ttcgtgatac ccctctttcc atgtattcac tacttctttc aaactctctc
     3661 tacgtttttt taattcttga ttttctgtgt aatagtctgt gctcttaata ttttcgtaat
     3721 catcaacaat ccgttcgaag agattgtttc ttgcaggcgt tcaaattcat cagcagttaa
     3781 tatctttcta ccagtctctt cacgtccaga gaacaaacct gtacgctcat tttcataatc
     3841 aaagggtttc gtagacctca tatgctctat tccactctgt aactgcttat ttgccttctg
     3901 taactcatcc ttaacttctt gcagttcctg tttatgaaat acagtatctt tcttgtactg
     3961 atccagttgc tcaaaaaaat ctcggtcaga tgttactagc aactcattta caagaacagc
     4021 atctttcctc gtttttcttg tacctgtttt ttgtgattca ataatttctt tgacacgttc
     4081 gttgtaatca atatttttat catttttcaa atcataattt tcacgtgttc gctcatggtc
     4141 aatatcatca ttcgttctac tttttcgctc tctttgatta tgaaattgca tgccttttag
     4201 tccagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4261 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4321 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
     4381 aattcgagct ccaattcgcc ctatagtgag tcgtattgac gtcgctattt aacgaccctg
     4441 ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta
     4501 tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat
     4561 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     4621 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     4681 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     4741 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     4801 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     4861 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     4921 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     4981 ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga
     5041 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     5101 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     5161 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     5221 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     5281 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa
     5341 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag
     5401 tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta
     5461 ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagctg
     5521 tccctcctgt tcagtactga cggggtggtc cgggagctgc atgtgtcaga ggttttcacc
     5581 gtcatcaccg aaacgacctt acgcctgttt aaactgcgaa ataaatctga catcgtttgt
     5641 atagaagtgg cggatatcat caatgccata tttcagcatc gcgatgcgct caacacccat
     5701 tccgaatccg aagccctgat attccttcgg atcaaagcca gccattttaa gcacgttcgg
     5761 gtgaaccatt ccggcaccga ggatttcaat ccagcctgtt cctttacata ctgagcagcc
     5821 gttcccaccg catttaaagc atgtcacatc cacttctact gaaggctcag taaacgggaa
     5881 gaagcttggg cggagtctga tttcacggtc ttgcccgaac atttttttcg caacaagttc
     5941 aagcgttcct tttaaatcac tcatgctgat gtttttgtca acgacaagcc cttcaatttg
     6001 cataaattgg tgagagtgcg tcgcatcatc gttatcacgg cgatatactt tacccgggca
     6061 aatgatttta acgggacctt tgccttcatg cttttccatc gtacgtgttt ggacaggaga
     6121 agtttgcgtt ctcatcaaag tttcctctgt gatgtaaaag ctgtcctgca tatcgcgcgc
     6181 tgggtgttct ttcggaagat tgagcgattc gaagttgtag taatccgttt caacctctgg
     6241 cccttcctcg actgtgtagc ccataccgat aaataaatct tcaatttctt caatgacaac
     6301 agtgagcgga tggcggccgc cgactgcaac agggttcccc ggcagcgtga cgtcaattgt
     6361 ctgtcctgca agcttctgtt tcatttcctc ttcttcaagc ttctcgtttt tgtcagcaat
     6421 cgcattggca atacgctccc ttacttcgtt cgcgagcgcc cccatttttg gacgttcctc
     6481 agcagaaagc ttgcccattc cgcgcagcac ttctgtaatc ggcccttttt ttccgagata
     6541 ttgcacccga atatcattga caaccttcaa tgagcttgcc gcttctactt gttctaaagc
     6601 ttcttgttcc agctgtttta gcttttcttc catctgcagc actctccttc tacatttttt
     6661 aacctaataa tgccaaatac cgtttgccac ccctctcttt tgataattat aatattggcg
     6721 aaattcgctt ctaaagatga aacgcaatat tatatgcttg ctttatcggc cgacacacat
     6781 tatgccacac cttgtagata aagtcaacaa ctttttgcaa aattgagctc gcgtaacggc
     6841 aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg
     6901 atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca
     6961 gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     7021 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     7081 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     7141 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     7201 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     7261 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     7321 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     7381 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     7441 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     7501 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     7561 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     7621 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     7681 atctattaat cagataaaat att
//
