LOCUS       sequence-210239-        5743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli/staphylococcal temperature-sensitive plasmid for allelic 
            exchange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5743)
  AUTHORS   Monk IR, Shah IM, Xu M, Tan MW, Foster TJ
  TITLE     Transforming the untransformable: application of direct 
            transformation to manipulate genetically Staphylococcus aureus and 
            Staphylococcus epidermidis.
  JOURNAL   MBio. 2012 Mar 20;3(2). pii: e00277-11. doi: 10.1128/mBio.00277-11. 
            Print 2012.
  PUBMED    22434850
REFERENCE   2  (bases 1 to 5743)
  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: "MBio."; 
            date: "2012-03-20"; volume: "3(2). pii: e00277-11. doi"; pages: " 
            10.1128/mBio.00277-11. Print 2012"
COMMENT     Sequence Label: pIMAY
FEATURES             Location/Qualifiers
     source          1..5743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(157..168)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2675..2693
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2706..2813
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2722..2738
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2723..2739
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2773..2789)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2773..2789)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2821..2840)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2822..2840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3213..3230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3347..3892)
                     /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(3427..3446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            4034..4143
                     /note="incP origin of transfer"
     protein_bind    4957..4975
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5042..5060
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5082..5708
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(5158..5177)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 gcatgcgttt tagcgtttat ttcgtttagt tatcggcata atcgttaaaa caggcgttat
       61 cgtagcgtaa aagcccttga gcgtagcgtg gctttgcagc gaagatgttg tctgttagat
      121 tatgaaagcc gatgactgaa tgaaataata agcgcagcgc ccttctattt cggttggagg
      181 aggctcaagg gagtatgagg gaatgaaatt ccctcatggg tttgatttta aaaattgctt
      241 gcaattttgc cgagcggtag cgctggaaaa tttttgaaaa aaatttggaa tttggaaaaa
      301 aatgggggga aaggaagcga attttgcttc cgtactacga ccccccatta agtgccgagt
      361 gccaattttt gtgccaaaaa cgctctatcc caactggctc aagggtttaa ggggtttttc
      421 aatcgccaac gaatcgccaa cgttttcgcc aacgtttttt ataaatctat atttaagtag
      481 ctttattgtt gtttttatga ttacaaagtg atacactaac tttataaaat tatttgattg
      541 gagtttttta aatggtgatt tcagaatcga aaaaaagagt tatgatttct ctgacaaaag
      601 agcaagataa aaaattaaca gatatggcga aacaaaaagg tttttcaaaa tctgcggttg
      661 cggcgttagc tatagaagaa tatgcaagaa aggaatcaga acaaaaaaaa taagcgaaag
      721 ctcgcgtttt tagaaggata cgagttttcg ctacttgttt ttgataaggt aattatatca
      781 tggctattaa aaatactaaa gctagaaatt ttggattttt attatatcct gactcaattc
      841 ctaatgattg gaaagaaaaa ttagagagtt tgggcgtatc tatggctgtc agtcctttac
      901 acgatatgga cgaaaaaaaa gataaagata catggaataa tagtaatatt atacaaaatg
      961 gaaagcacta taaaaaacca cactatcacg ttatatatat tgcacgaaat cctgtaacaa
     1021 tagaaagcgt taggaacaag attaagcgaa aattggggaa tagttcagtt gctcatgttg
     1081 agatacttga ttatatcaaa ggttcatatg aatatttgac tcatgaatca aaggacgcta
     1141 ttgctaagaa taaacatata tacgacaaaa aagatatttt gaacattaat gattttgata
     1201 ttgaccgcta tataacactt gatgaaagcc aaaaaagaga attgaagaat ttacttttag
     1261 atatagtgga tgactataat ttggtaaata caaaagattt aatggctttt attcgcctta
     1321 ggggagcgga gtttggaatt ttaaatacga atgatgtaaa agatattgtt tcaacaaact
     1381 ctagcgcctt tagattatgg tttgagggca attatcagtg tggatataga gcaagttatg
     1441 caaaggttct tgatgctgaa acgggggaaa taaaatgaca aacaaagaaa aagagttatt
     1501 tgctgaaaat gaggaattaa aaaaagaaat taaggactta aaagagcgta ttgaaagata
     1561 cagagaaatg gaagttgaat taagtacaac aatagattta ttgagaggag ggattattga
     1621 ataaataaaa gccccctgac gaaagtcgaa gggggttttt attttggttt gatgttgcga
     1681 ttaatagcaa tacattctat aatagaaggt atggaggatg ttatataatg agacagaatt
     1741 atgatgatca tatgtcaact aacggggcag gttagtgaca ttagaaaacc gactgtaaaa
     1801 agtacagtcg gcattatctc atattataaa agccagtcat taggcctatc tgacaattcc
     1861 tgaatagagt tcataaacaa tcctgcatga taaccatcac aaacagaatg atgtacctgt
     1921 aaagatagcg gtaaatatat tgaattacct ttattaatga attttcctgc tgtaataatg
     1981 ggtagaaggt aattactatt attattgata tttaagttaa acccagtaaa tgaagtccat
     2041 ggaataatag aaagagaaaa agcattttca ggtataggtg ttttgggaaa caatttcccc
     2101 gaaccattat atttctctac atcagaaagg tataaatcat aaaactcttt gaagtcattc
     2161 tttacaggag tccaaatacc agagaatgtt ttagatacac catcaaaaat tgtataaagt
     2221 ggctctaact tatcccaata acctaactct ccgtcgctat tgtaaccagt tctaaaagct
     2281 gtatttgagt ttatcaccct tgtcactaag aaaataaatg cagggtaaaa tttatatcct
     2341 tcttgtttta tgtttcggta taaaacacta atatcaattt ctgtggttat actaaaagtc
     2401 gtttgttggt tcaaataatg attaaatatc tcttttctct tccaattgtc taaatcaatt
     2461 ttattaaagt tcatgggttt cactctcctt ctacattttt taacctaata atgccaaata
     2521 ccgtttgcca cccctctctt tgataattat aatattggcg aaattcgctt ctaaagatga
     2581 aacgcaatat tatatgcttg ctttatcggc cgtatgtgat tataccagcc ccctcactac
     2641 atgtcaagaa taaactgcca aagcataatg ggataattaa ccctcactaa agggaacaaa
     2701 agctgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc
     2761 tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag ctccaattcg
     2821 ccctatagtg agtcgtatta cgacgtccca gggcttcccg gtatcaacag ggacaccagg
     2881 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc
     2941 gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg
     3001 cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca
     3061 aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga
     3121 tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag
     3181 cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     3241 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     3301 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     3361 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     3421 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     3481 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     3541 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     3601 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     3661 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     3721 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     3781 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     3841 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     3901 tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc
     3961 acctgaagtc agccccatac gatataagtt gtaattctcc gccgcttgcc ctcatctgtt
     4021 acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc
     4081 gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg
     4141 cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct
     4201 gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag
     4261 ggttatgcag cggaaaagcg ctgcttccct gctgttttgt ggaatatcta ccgactggaa
     4321 acaggcaaat gcaggaaatt actgaactga ggggacaggc gagaggagat cttgatctaa
     4381 tgattcaaac ccttgtgaac ttctttagaa caaaagaggt tcgtaacaag attttcttca
     4441 cactagcaat gttagtaatt tttaaaatag ggacttatat accagctcca ggagtaaatc
     4501 ctgcagcttt tgataatccc caaggttctc aaggtgccac tgagttatta aatacttttg
     4561 gtggcggagc cttgaaacga ttttctattt ttgcaatggg tattgtaccc tacatcactg
     4621 catcaatcgt aatgcaatta ttacaaatgg atattgtccc taaattctca gaatgggcaa
     4681 aacaaggtga agtaggtaga agaaagttaa ataacgttac tcgttattta gcaatttctt
     4741 tagcatttat ccaatctata ggtatggcat tccaatttaa taattatctc aaaggtgcgc
     4801 tgattatcaa tcagtcaatt atgagttatt tattaatagc actagttttg acagcaggaa
     4861 ctgctttctt aatatggctt ggtgatcaaa tcactcagtt cggtgttggt aatggtattt
     4921 ctattatcat attcccatca agcttatttt aattatactc tatcaatgat agagtgtcaa
     4981 tatttttttt agtttttcat gaactcgagg ggatccaaat aaaaaactag tttgacaaat
     5041 aactctatca atgatagagt gtcaacaaaa aggaggaatt aatgatgtct agattagata
     5101 aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa
     5161 caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa
     5221 aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc
     5281 acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt
     5341 ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta
     5401 cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt
     5461 cactagagaa tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat
     5521 tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta
     5581 tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct
     5641 tcttattcgg ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg
     5701 ggtcttaaaa gcagcataac ctttttccgt gatggtaact tca
//
