LOCUS       sequence-332204-        6411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes an IPTG inducible recT from E. faecium Com15 used to 
            ectopically express recT in Enterococcus for recombineering..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  AUTHORS   Chen V, Griffin ME, Maguin P, Varble A, Hang HC
  TITLE     RecT recombinase expression enables efficient gene editing in 
            Enterococcus.
  JOURNAL   Appl Environ Microbiol. 2021 Jul 7:AEM0084421. doi: 
            10.1128/AEM.00844-21.
  PUBMED    34232061
REFERENCE   2  (bases 1 to 6411)
  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. 2021 Jul 7:AEM0084421. doi: 
            10.1128/AEM.00844-21."
COMMENT     Sequence Label: pRecT_2
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2246..2332
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2351..2378
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2458..2492
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    2596..2612
                     /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)."
     CDS             3862..4944
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3881..3900)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(5922..5933)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aaaatctgcg gttgcggcgt tagctataga agaatatgca agaaaggaat cagaacaaaa
       61 aaaataagcg aaagctcgcg tttttagaag gatacgagtt ttcgctactt gtttttgata
      121 aggtaattat atcatggcta ttaaaaatac taaagctaga aattttggat ttttattata
      181 tcctgactca attcctaatg attggaaaga aaaattagag agtttgggcg tatctatggc
      241 tgtcagtcct ttacacgata tggacgaaaa aaaagataaa gatacatgga atagtagtga
      301 tgttatacga aatggaaagc actataaaaa accacactat cacgttatat atattgcacg
      361 aaatcctgta acaatagaaa gcgttaggaa caagattaag cgaaaattgg ggaatagttc
      421 agttgctcat gttgagatac ttgattatat caaaggttca tatgaatatt tgactcatga
      481 atcaaaggac gctattgcta agaataaaca tatatacgac aaaaaagata ttttgaacat
      541 taatgatttt gatattgacc gctatataac acttgatgaa agccaaaaaa gaaaattgaa
      601 gaatttactt ttagatatag tggatgacta taatttggta aatacaaaag atttaatggc
      661 ttttattcgc cttaggggag cggagtttgg aattttaaat acgaatgatg taaaagatat
      721 tgtttcaaca aactctagcg cctttagatt atggtttgag ggcaattatc agtgtggata
      781 tagagcaagt tatgcaaagg ttcttgatgc tgaaacgggg gaaataaaat gacaaacaaa
      841 gaaaaagagt tatttgctga aaatgaggaa ttaaaaaaag aaattaagga cttaaaagag
      901 cgtattgaaa gatacagaga aatggaagtt gaattaagta caacaataga tttattgaga
      961 ggagggatta ttgaataaat aaaagccccc ctgacgaaag tccaagggtt tattgttttc
     1021 taaaatctga ttaccaatta gaatgaatat ttcccaaata ttaaataata aaacaaaaaa
     1081 attaaaaaag tgtttccacc attttttcaa tttttttata atttttttaa tctgttattt
     1141 aaatagttta tagttaaatt tacattttca ttagtccatt caatattctc tccaagataa
     1201 ctacgaactg ctaacaaaat tctctcccta tgttctaatg gagaagattc agccactgca
     1261 tttcccgcaa tatcttttgg tatgatttta cccgtgtcca tagttaaaat catacggcat
     1321 aaagttaata tagagttggt ttcatcatcc tgataattat ctattaattc ctctgacgaa
     1381 tccataatgg ctcttctcac atcagaaaat ggaatatcag gtagtaattc ctctaagtca
     1441 taatttccgt atattctttt attttttcgt tttgcttggt aaagcattat ggttaaatct
     1501 gaatttaatt ccttctgagg aatgtatcct tgttcataaa gctcttgtaa ccattctcca
     1561 taaataaatt cttgtttggg aggatgattc cacggtacca tttcttgctg aataataatt
     1621 gttaattcaa tatatcgtaa gttgctttta tctcctattt tttttgaaat aggtctaatt
     1681 ttttgtataa gtatttcttt actttgatct gtcaatggtt cagatacgac gactaaaaag
     1741 tcaagatcac tatttggttt tagtccactc tcaactcctg atccaaacat gtaagtacca
     1801 ataaggttat tttttaaatg tttccgaagt atttttttca ctttattaat ttgttcgtat
     1861 gtattcaaat atatcctcct cactattttg attagtacct attttatatc catagttgtt
     1921 aattaaataa acttaattta gtttatttat agatttcatt ggcttctaaa ttttttatct
     1981 agataataat tattttagtt aattttattc tagattatat atgatatgat ctttcatttc
     2041 cataaaacta aagtaagtgt aaacctattc attgttttaa aaatatctct tgccagtcac
     2101 gttacgttat tagttatagt tattataaca tgtattcacg aacgaaaatc gacctgcagg
     2161 catgcaagct tggcgttata aaattagtat aattatagca cgagctctga taaatatgaa
     2221 catgcgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2281 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgct
     2341 ctagctaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttgtt
     2401 aactctagag ctgcctgccg cgtttcggtg atgaagatct tcccgatgat taattaattc
     2461 agaacgctcg gttgccgccg ggcgtttttt atgcagcaat ggcaagaacg ttgctcgagg
     2521 gtaaatgtga gcactcacaa ttcattttgc aaaagttgtt gactttatct acaaggtgtg
     2581 gcataatgtg tggaattgtg agcggataac aattaagctt gtacttagga ggatgattat
     2641 ttatggcaac aaatgaatcg ttaaaaaacc aattggcaga aaagccacag aaacaagttg
     2701 caccaggaca gttagggctt aaagctctaa tgaatacacc aacaatgaga aagaaatttg
     2761 aagaagtact tcatgacaat gctaatgctt ttatgtcgaa tgttatgact cttgtatcta
     2821 atgacaatta tcttgcagat agtgaaccga tgtctatcat gagtggtgcg ttaactgctg
     2881 caacattaaa tcttggacta gataagaatc ttggctatgc ctatctcgtt ccatttaata
     2941 gcaaaaacaa gcaaacagga aaatgggaaa agaaagctca atttatgctt ggctataaag
     3001 gatatatcca attagcccaa cgatcaggta aatacaaagc attaaatgtg attgaagttt
     3061 acgaaggaga actaaaaagc tggaaccgac tgacagaaga gttcgagttt gatccaaatg
     3121 gtagaacatc tgatgaagtc attggatatg ttggctattt tgaattactg aatggattca
     3181 agaaaactgt ctattggacc aaacaagaaa ttgaagctca tcgaattggt aacaataaag
     3241 atcgagataa gacaaagtta agtggtgtgt gggcatctga ttacaatgca atggcacgaa
     3301 aaactgtttt gagaaatctt ctttctaaat gggggatctt atccattgaa atgcaagaag
     3361 ctaccacatc ggatgagaga gtccaaagag ttcaagaaga tggcagcatt attgctgaaa
     3421 cagaagttga ggaagatatt cctgaaagaa aagaagcaga aaaagtagaa ataaacgctg
     3481 acaatggatc aacagctttt gaagagtatg cagaatcagt tcagacagat gaagcacaag
     3541 aatcgctgtt cgatgcatcg aatccaccat taaataaata gcccgggtaa ctagtaactg
     3601 cagatgctag ctcgcatgca agctaattcg gtggaaacga ggtcatcatt tccttccgaa
     3661 aaaacggttg catttaaatc ttacatatgt aatactttca aagactacat ttgtaagatt
     3721 tgatgtttga gtcggctgaa agatcgtacg taccaattat tgtttcgtga ttgttcaagc
     3781 cataacactg tagggatagt ggaaagagtg cttcatctgg ttacgatcaa tcaaatattc
     3841 aaacggaggg agacgatttt gatgaaacca gtaacgttat acgatgtcgc agagtatgcc
     3901 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     3961 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     4021 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     4081 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     4141 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaaacggc ggtgcacaat
     4201 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     4261 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     4321 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     4381 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     4441 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     4501 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     4561 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     4621 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     4681 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     4741 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     4801 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     4861 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4921 tcccgactgg aaagcgggca gtgaatgagt gatcgttaaa tttatactgc aatcggatgc
     4981 gattattgaa taaaagatat gagagattta tctaatttct tttttcttgt aaaaaaagaa
     5041 agttcttaaa ggttttatag ttttggtcgt aggaagatct ggatcctaat gaattcatct
     5101 gcaggggccc atcagtctga cgaccaagag agccataaac accaatagcc ttaacatcat
     5161 ccccatattt atccaatatt cgttccttaa tttcatgaac aatcttcatt ctttcttctc
     5221 tagtcattat tattggtcca ttcactattc tcattccctt ttcagataat tttagatttg
     5281 cttttctaaa taagaatatt tggagagcac cgttcttatt cagctattaa taactcgtct
     5341 tcctaagcat ccttcaatcc ttttaataac aattatagca tctaatcttc aacaaactgg
     5401 cccgtttgtt gaactactct ttaataaaat aatttttccg ttcccaattc cacattgcaa
     5461 taatagaaaa tccatcttca tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt
     5521 cttctgtgtc atcaaggttt aattttttat gtatttcttt taacaaacca ccataggaga
     5581 ttaacctttt acggtgtaaa ccttcctcca aatcagacaa acgtttcaaa ttcttttctt
     5641 catcatcggt cataaaatcc gtatccttta caggatattt tgcagtttcg tcaattgccg
     5701 attgtatatc cgatttatat ttatttttcg gtcgattttt tattaaaacg tctcaaaatc
     5761 gtttctgaga cgttttagcg tttatttcgt ttagttatcg gcataatcgt taaaacaggc
     5821 gttatcgtag cgtaaaagcc cttgagcgta gcgtggcttt gcagcgaaga tgttgtctgt
     5881 tagattatga aagccgatga ctgaatgaaa taataagcgc agcgtccttc tatttcggtt
     5941 ggaggaggct caagggagtt tgagggaatg aaattccctc atgggtttga ttttaaaaat
     6001 tgcttgcaat tttgccgagc ggtagcgctg gaaaattttt gaaaaaaatt tggaatttgg
     6061 aaaaaaatgg ggggaaagga agcgaatttt gcttccgtac tacgaccccc cattaagtgc
     6121 cgagtgccaa tttttgtgcc aaaaacgctc tatcccaact ggctcaaggg tttgaggggt
     6181 ttttcaatcg ccaacgaatc gccaacgttt tcgccaacgt tttttataaa tctatattta
     6241 agtagcttta ttgttgtttt tatgattaca aagtgataca ctaattttat aaaattattt
     6301 gattggagtt ttttaaatgg tgatttcaga atcgaaaaaa agagttatga tttctctgac
     6361 aaaagagcaa gataaaaaat taacagatat ggcgaaacaa aaaggttttt c
//
