LOCUS       sequence-458137-        8136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing RecE*T controled by Ptrc promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8136)
  AUTHORS   Wang X, Yang S, Qian F, Dong F, Zhou X, Yin M, Zhang Y, Huang Z, 
            Jiang Y, Yang S
  TITLE     RAGATH-Associated DNA Nuclease Assisted DNA Insertion in 
            Corynebacterium glutamicum.
  JOURNAL   ACS Synth Biol. 2025 May 8. doi: 10.1021/acssynbio.5c00022.
  PUBMED    40338137
REFERENCE   2  (bases 1 to 8136)
  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: "ACS Synth 
            Biol. 2025 May 8. doi: 10.1021/acssynbio.5c00022."
COMMENT     Sequence Label: pRecE564T
FEATURES             Location/Qualifiers
     source          1..8136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3495..4577)
                     /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
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4539..4558
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4578..4655)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4861..4880
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    5021..5037
                     /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     5026..5048
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(7301..7318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7472..8060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7552..7571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actaataacg taacgtgact ggcaagagat atttttaaaa caatgaatag gtttacactt
       61 actttagttt tatggaaatg aaagatcata tcatatataa tctagaataa aattaactaa
      121 aataattatt atctagataa aaaatttaga agccaatgaa atctataaat aaactaaatt
      181 aagtttattt aattaacaac tatggatata aaataggtac taatcaaaat agtgaggagg
      241 atatatttga atacatacga acaaattaat aaagtgaaaa aaatacttcg gaaacattta
      301 aaaaataacc ttattggtac ttacatgttt ggatcaggag ttgagagtgg actaaaacca
      361 aatagtgatc ttgacttttt agtcgtcgta tctgaaccat tgacagatca aagtaaagaa
      421 atacttatac aaaaaattag acctatttca aaaaaaatag gagataaaag caacttacga
      481 tatattgaat taacaattat tattcagcaa gaaatggtac cgtggaatca tcctcccaaa
      541 caagaattta tttatggaga atggttacaa gagctttatg aacaaggata cattcctcag
      601 aaggaattaa attcagattt aaccataatg ctttaccaag caaaacgaaa aaataaaaga
      661 atatacggaa attatgactt agaggaatta ctacctgata ttccattttc tgatgtgaga
      721 agagccatta tggattcgtc agaggaatta atagataatt atcaggatga tgaaaccaac
      781 tctatattaa ctttatgccg tatgatttta actatggaca cgggtaaaat cataccaaaa
      841 gatattgcgg gaaatgcagt ggctgaatct tctccattag aacataggga gagaattttg
      901 ttagcagttc gtagttatct tggagagaat attgaatgga ctaatgaaaa tgtaaattta
      961 actataaact atttaaataa cagattaaaa aaattataaa aaaattgaaa aaatggtgga
     1021 aacacttttt tcaatttttt tgttttatta tttaatattt gggaaatatt cattctaatt
     1081 ggtaatcaga ttttagaaaa caataaaccc ttgcatatga tatcgatgta cagatctttt
     1141 tttaaggcag ttattggtgc ccttcgaaat gaccgaccaa gcgacgccca acctgccatc
     1201 acgagatttc gattccaccg ccgccttcta tgaaagccat caatcctgcc tatttgccac
     1261 gtttaacaag gtagttaagc gttcatttac gaagaaaaca cgataagctg cacaaatacc
     1321 tgaaaaagtt gaacgccccg tgagcgggaa ctcacagggc gtcggctaac ccccagtcat
     1381 cagctgggag aaagcactca agacatgact ctagccgatc cgcaggacac agtcacagct
     1441 agcgcgtgga aattttccgc cgatctgttc gacacccacc ccgaactagc gctgcgctca
     1501 cgcggctgga cggcagaaga tcgccgcgaa ctgctcgctc acctgggacg cgaaagcttc
     1561 cagggcagca agacaagaga tttcgcgagc gcctggatta aaaacccgga taccggcgaa
     1621 acccaaccaa agctctaccg ggctggctca aaagcgctga cgcggtgcca gtacgttgcg
     1681 ctgacgcacg cgcaacatgc cgcggtgatc gtgcttgaca tcgatgtgcc cagccaccag
     1741 gccggcggga agattgagca cgtaaacccg caggtctacg cgattttaga gaaatgggca
     1801 cgcctagaaa aagcgccggc ttggatcggc gtgaatccgc tgagcgggaa atgccagctc
     1861 atctggctca ttgacccggt gtatgccgca gcaggtaaaa ccagcccaaa tatgcgcctg
     1921 ctggctgcaa cgacggaaga aatgactcgt gttttcggcg ctgaccaggc tttttcgcat
     1981 aggctgagcc ggtggccgct gcacgtctca gacgatccga cagcctataa atggcactgc
     2041 cagcatgatc gtgtggatcg gctggccgac ctaatggaga ttgctcgaac gatgaccgga
     2101 tcacagaagc cgaaaaagta cattgagcag gacttttcca gcggacgcgc ccgcattgaa
     2161 gcggcacaac gcgccaccgc agaagccaag gcgctagcga ttttggacgc gagcctgccg
     2221 agcgccctgg acgcgtccgg cgacctgatc gacggcgtgc gagtgctctg gacaaatcca
     2281 gagcgcgcag cgcgcgacga gaccgcgttt cgccacgcgt tgaccgtggg ataccagctc
     2341 aaagctgctg gtgagcgcct aaaagatgcc aagatcatcg acgcgtatga agtggcgtac
     2401 aacgttgccc aggcggtcgg tgcagacggc cgggagccgg atcttcccgc catgcgtgat
     2461 cgcctgacga tggcgcgtcg tgtgcgcggc tacgtggcta aaggccagcc agtcgtccct
     2521 gctcgtcggg tggaaacgca gagcagccga gggcggaaag ctctagcgac gatggggcga
     2581 cggggcgcag ctacatcgaa tgcacgcaga tgggctgacc cagaaagtaa gtatgcgcag
     2641 gagacgcgac agcgattagc ggaagcaaac aaacgccgag aaatgacagg cgagttgctc
     2701 gaacttcgcg tcaaaactgc gatcctggat gcccgttctc aatcggttgc tgatccctcg
     2761 actcgtgagc ttgcaggcga actaggtgtc agtgaaaggc gcatccaaca agtcagaaag
     2821 gcacttggaa tggaagctaa acgcggccgt ccacgggctg aaaactaata aacgaaacac
     2881 cgtcagcaga aaacggttcc cccctttagg ggtcccgtcc ttgctctggc tctcacttgc
     2941 cctcaccctc cgctatccac gggctgaaaa ctaataaacg aaacaccgtc agcagaaaac
     3001 ggttcccccc ctttagggtg tctcgctcct agctctgatc cctccccggt tcctccccgg
     3061 cctgattttt aagggggctc acgctgtcgg cagagaacgg ttcccccgcc ttctgctctg
     3121 gctcttcctc gactccctcc ccctcaaaaa tctcctcgag tctagagtag tgggcagtgt
     3181 gcacaccata gtgggcatga gtaataccta cgcgcgcgtg ggctagggct taacgcgcgt
     3241 tttgccgtgc tgcggggcat acgttagcgc atacgctttt ttctgtgaaa cctttttgtg
     3301 ttgttgtttc gtgttggttt cctttctgtt ggcggggcaa cttaacgcct gcgggggtgg
     3361 ttgttgacgt taacggtgcc tatgagatct tccggaatcg atggaaggag ctgactgggt
     3421 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     3601 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     3661 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     3721 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     3781 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     3841 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     3901 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     3961 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4021 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4081 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4141 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4201 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4261 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4321 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4381 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4441 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4501 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     4561 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     4621 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     4681 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     4741 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     4801 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     4861 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     4921 gatgcgtccg gcgtagagga tcgagataag cttcggttct ggcaaatatt ctgaaatgag
     4981 ctgttgacat tgtgagcgct cacaattata atgtgtggaa ttgtgagcgg ataacaattt
     5041 cacacaggat ctagagctag ctaatctgag ttgaggttaa aaaacaatgg aacatccgca
     5101 caatgagaat gctggcagcg atccgcatcg cgattgcagt gatgaaactg gcgaagtcgc
     5161 agatcccgta atcgtagaag acatagagcc aggtatttat tacggaattt cgaatgagaa
     5221 ttaccacgcg ggtcccggta tcagtaagtc tcagctcgat gacattgctg atactccggc
     5281 actatatttg tggcgtaaaa atgcccccgt ggacaccaca aagacaaaaa cgctcgattt
     5341 aggaactgct ttccactgcc gggtacttga accggaagaa ttcagtaacc gctttatcgt
     5401 agcacctgaa tttaaccgcc gtacaaacgc cggaaaagaa gaagagaaag cgtttctgat
     5461 ggaatgcgca agcacaggaa aaacggttat cactgcggaa gaaggccgga aaattgaact
     5521 catgtatcaa agcgttatgg ctttgccgct ggggcaatgg cttgttgaaa gcgccggaca
     5581 cgctgaatca tcaatttact gggaagatcc tgaaacagga attttgtgtc ggtgccgtcc
     5641 ggacaaaatt atccctgaat ttcactggat catggacgtg aaaactacgg cggatattca
     5701 acgattcaaa accgcttatt acgactaccg ctatcacgtt caggatgcat tctacagtga
     5761 cggttatgaa gcacagtttg gagtgcagcc aactttcgtt tttctggttg ccagcacaac
     5821 tattgaatgc ggacgttatc cggttgaaat tttcatgatg ggcgaagaag caaaactggc
     5881 aggtcaacag gaatatcacc gcaatctgcg aaccctgtct gactgcctga ataccgatga
     5941 atggccagct attaagacat tatcactgcc ccgctgggct aaggaatatg caaatgacta
     6001 agcaaccacc aatcgcaaaa gccgatctgc aaaaaactca gggaaaccgt gcaccagcag
     6061 cagttaaaaa tagcgacgtg attagtttta ttaaccagcc atcaatgaaa gagcaactgg
     6121 cagcagctct tccacgccat atgacggctg aacgtatgat ccgtatcgcc accacagaaa
     6181 ttcgtaaagt tccggcgtta ggaaactgtg acactatgag ttttgtcagt gcgatcgtac
     6241 agtgttcaca gctcggactt gagccaggta gcgccctcgg tcatgcatat ttactgcctt
     6301 ttggtaataa aaacgaaaag agcggtaaaa agaacgttca gctaatcatt ggctatcgcg
     6361 gcatgattga tctggctcgc cgttctggtc aaatcgccag cctgtcagcc cgtgttgtcc
     6421 gtgaaggtga cgagtttagc ttcgaatttg gccttgatga aaagttaata caccgcccgg
     6481 gagaaaacga agatgccccg gttacccacg tctatgctgt cgcaagactg aaagacggag
     6541 gtactcagtt tgaagttatg acgcgcaaac agattgagct ggtgcgcagc ctgagtaaag
     6601 ctggtaataa cgggccgtgg gtaactcact gggaagaaat ggcaaagaaa acggctattc
     6661 gtcgcctgtt caaatatttg cccgtatcaa ttgagatcca gcgtgcagta tcaatggatg
     6721 aaaaggaacc actgacaatc gatcctgcag attcctctgt attaaccggg gaatacagtg
     6781 taatcgataa ttcagaggaa taattcagcc tggcggtgta atgcccatgg actatcgcta
     6841 atcgtaatcg cggagatcca gagtaacaaa aataacgccc caccttgata atgaggtggg
     6901 gcgttatttt tgtgtcttaa tccagtaggt agtctgccaa gatcaggtta gtgcagtacc
     6961 aattgttgtt cttggagaag ttaacagcct ctgccatact caacgcaatg tccttatcac
     7021 tggtcttagt gttcacgtac cgtacccggc ctgtggtcat ttcgatctgt ctcataaccc
     7081 aattctctgg atccgggacc gagctcgaat tacgtcgcgg tgagttcagg ctttttcata
     7141 tctcattgcc cccggacgag cgtctgctcc gccattcgcc gtccgccgtg ccaatcggat
     7201 cagccgtcca atccttgggc cacgctaagc ttcatatgcg gtgtgaaata ccgcacagat
     7261 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     7321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8101 ggaacgaaaa ctcacgttaa gggattttgg tcatga
//
