LOCUS       sequence-71047-e        8185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  I-SceI DSB repair reporter cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8185)
  AUTHORS   Gupta R, Barkan D, Redelman-Sidi G, Shuman S, Glickman MS
  TITLE     Mycobacteria exploit three genetically distinct DNA double-strand 
            break repair pathways.
  JOURNAL   Mol Microbiol. 2011 Jan;79(2):316-30. doi: 
            10.1111/j.1365-2958.2010.07463.x. Epub 2010 Nov 24.
  PUBMED    21219454
REFERENCE   2  (bases 1 to 8185)
  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: 
            "10.1111/j.1365-2958.2010.07463.x"; journalName: "Mol Microbiol"; 
            date: "2011-01"; volume: "79"; issue: "2"; pages: "316-30"
COMMENT     Sequence Label: pRGM10
FEATURES             Location/Qualifiers
     source          1..8185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(17..22,23..40,41..47)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    53..72
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     complement(173..195)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1497..1543
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2741..3556
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3888..4476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4377..4396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      7546..7592
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gatccagacc ccaggcttga cactttatgc ttccggctcg tataatgtgt ggaattgtga
       61 gcgctcacaa ttcgcatgcg gtaccaagct tgatccgata acacaggaac agatctatgg
      121 ttcgtgcaaa caaacgcaac gaggctctac gaatcggaag cttcgatccc gtcgttttac
      181 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      241 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      301 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
      361 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
      421 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
      481 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
      541 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttaactcg gcgtttcatc
      601 tgtggtgcaa cgggcgctgg gtcggttacg gccaggacag tcgtttgccg tctgaatttg
      661 acctgagcgc atttttacgc gccggagaaa accgcctcgc ggtgatggtg ctgcgctgga
      721 gtgacggcag ttatctggaa gatcaggata tgtggcggat gagcggcatt ttccgtgacg
      781 tctcgttgct gcataaaccg actacacaaa tcagcgattt ccatgttgcc actcgcttta
      841 atgatgattt cagccgcgct gtactggagg ctgaagttca gatgtgcggc gagttgcgtg
      901 actacctacg ggtaacagtt tctttatggc agggtgaaac gcaggtcgcc agcggcaccg
      961 cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta tgccgatcgc gtcacactac
     1021 gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat cccgaatctc tatcgtgcgg
     1081 tggttgaact gcacaccgcc gacggcacgc tgattgaagc agaagcctgc gatgtcggtt
     1141 tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa cggcaagccg ttgctgattc
     1201 gaggcgttaa ccgtcacgag catcatcctc tgcatggtca ggtcatggat gagcagacga
     1261 tggtgcagga tatcctgctg atgaagcaga acaactttaa cgccgtgcgc tgttcgcatt
     1321 atccgaacca tccgctgtgg tacacgatat tatcctgtta tccctaccat ggtagggata
     1381 acagggtaat agctcctgaa atgaattcgg ggatcagctg cagaattatt accctgttat
     1441 ccctaagctt atcgatgtcg acgtagttaa ctagcgtacg atcgactgcc aggcatcaaa
     1501 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatgccatca tggccgcggt
     1561 ggagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg gtgaagtgcc
     1621 tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac cgcagccgga
     1681 gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga ccgcatggtc
     1741 agaagccggg cacatcagcg cctggcagca gtggcgtctg gcggaaaacc tcagtgtgac
     1801 gctccccgcc gcgtcccacg ccatcccgca tctgaccacc agcgaaatgg atttttgcat
     1861 cgagctgggt aataagcgtt ggcaatttaa ccgccagtca ggctttcttt cacagatgtg
     1921 gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat cagttcaccc gtgcaccgct
     1981 ggataacgac attggcgtaa gtgaagcgac ccgcattgac cctaacgcct gggtcgaacg
     2041 ctggaaggcg gcgggccatt accaggccga agcagcgttg ttgcagtgca cggcagatac
     2101 acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg cagcatcagg ggaaaacctt
     2161 atttatcagc cggaaaacct accggattga tggtagtggt caaatggcga ttaccgttga
     2221 tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt ggcctgaact gccagctggc
     2281 gcaggtagca gagcgggtaa actggctcgg attagggccg caagaaaact atcccgaccg
     2341 ccttactgcc gcctgttttg accgctggga tctgccattg tcagacatgt ataccccgta
     2401 cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc gaattgaatt atggcccaca
     2461 ccagtggcgc ggcgacttcc agttcaacat cagccgctac agtcaacagc aactgatgga
     2521 aaccagccat cgccatctgc tgcacgcgga agaaggcaca tggctgaata tcgacggttt
     2581 ccatatgggg attggtggcg acgactcctg gagcccgtca gtatcggcgg aattccagct
     2641 gacgccggtc gcctaccatt accagttggt ctggtgtcaa aaataataat catgaacaat
     2701 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2761 aacgtcttgc tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2821 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2881 ccatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2941 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     3001 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
     3061 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3121 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3181 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3241 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata atcttttgcc
     3301 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3361 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3421 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3481 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3541 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3601 gacttgacgg gacggcggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     3661 atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc
     3721 aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat
     3781 gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg cagacctcac
     3841 tagttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4021 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4261 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4501 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4561 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4621 accgagcgca acgcgtgcgg ccgcggtacc agatctttgt ctcgttgctg cataaaccga
     4681 ctacacaaat cagcgatttc catgttgcca ctcgctttaa tgatgatttc agccgcgctg
     4741 tactggaggc tgaagttcag atgtgcggcg agttgcgtga ctacctacgg gtaacagttt
     4801 ctttatggca gggtgaaacg caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta
     4861 tcgatgagcg tggtggttat gccgatcgcg tcacactacg tctgaacgtc gaaaacccga
     4921 aactgtggag cgccgaaatc ccgaatctct atcgtgcggt ggttgaactg cacaccgccg
     4981 acggcacgct gattgaagca gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa
     5041 atggtctgct gctgctgaac ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc
     5101 atcatcctct gcatggtcag gtcatggatg agcagacgat ggtgcaggat atcctgctga
     5161 tgaagcagaa caactttaac gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt
     5221 acacgctgtg cgaccgctac ggcctgtatg tggtggatga agccaatatt gaaacccacg
     5281 gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg atgagcgaac
     5341 gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg
     5401 ggaatgaatc aggccacggc gctaatcacg acgcgctgta tcgctggatc aaatctgtcg
     5461 atccttcccg cccggtgcag tatgaaggcg gcggagccga caccacggcc accgatatta
     5521 tttgcccgat gtacgcgcgc gtggatgaag accagccctt cccggctgtg ccgaaatggt
     5581 ccatcaaaaa atggctttcg ctacctggag agacgcgccc gctgatcctt tgcgaatacg
     5641 cccacgcgat gggtaacagt cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt
     5701 atccccgttt acagggcggc ttcgtctggg actgggtgga tcagtcgctg attaaatatg
     5761 atgaaaacgg caacccgtgg tcggcttacg gcggtgattt tggcgatacg ccgaacgatc
     5821 gccagttctg tatgaacggt ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg
     5881 aagcaaaaca ccagcagcag tttttccagt tccgtttatc cgggcaaacc atcgaagtga
     5941 ccagcgaata cctgttccgt catagcgata acgagctcct gcactggatg gtggcgctgg
     6001 atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt
     6061 tgattgaact gcctgaacta ccgcagccgg agagcgccgg gcaactctgg ctcacagtac
     6121 gcgtagtgca accgaacgcg accgcatggt cagaagccgg gcacatcagc gcctggcagc
     6181 agtggcgtct ggcggaaaac ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc
     6241 atctgaccac cagcgaaatg gatttttgca tcgagctggg taataagcgt tggcaattta
     6301 accgccagtc aggctttctt tcacagatgt ggattggcga taaaaaacaa ctgctgacgc
     6361 cgctgcgcga tcagttcacc cgtgcaccgc tggataacga cattggcgta agtgaagcga
     6421 cccgcattga ccctaacgcc tgggtcgaac gctggaaggc ggcgggccat taccaggccg
     6481 aagcagcgtt gttgcagtgc acggcagata cacttgctga tgcggtgctg attacgaccg
     6541 ctcacgcgtg gcagcatcag gggaaaacct tatttatcag ccggaaaacc taccggattg
     6601 atggtagtgg tcaaatggcg attaccgttg atgttgaagt ggcgagcgat acaccgcatc
     6661 cggcgcggat tggcctgaac tgccagctgg cgcaggtagc agagcgggta aactggctcg
     6721 gattagggcc gcaagaaaac tatcccgacc gccttactgc cgcctgtttt gaccgctggg
     6781 atctgccatt gtcagacatg tataccccgt acgtcttccc gagcgaaaac ggtctgcgct
     6841 gcgggacgcg cgaattgaat tatggcccac accagtggcg cggcgacttc cagttcaaca
     6901 tcagccgcta cagtcaacag caactgatgg aaaccagcca tcgccatctg ctgcacgcgg
     6961 aagaaggcac atggctgaat atcgacggtt tccatatggg gattggtggc gacgactcct
     7021 ggagcccgtc agtatcggcg gaattccagc tgacgccggt cgcctaccat taccagttgg
     7081 tctggtgtca aaaataataa tcatgaacaa taaaactgtc tgcttacata aacagtaata
     7141 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt
     7201 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     7261 gtgcgacaat ctatcgcttg tatgggaagc cccatgcgcc agagttgttt ctgaaacatg
     7321 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     7381 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     7441 tcaccactgc gatccccaaa tctagatatc catggatcca gctgcagaat tcgaagctta
     7501 tcgatgtcga cgtagttaac tagcgtacga tcgactgcca ggcatcaaat aaaacgaaag
     7561 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtccggcca tcatggccgc
     7621 ggtgatcagc tagagccgtg aacgacaggg cgaacgccag cccgccgacg gcgagggttc
     7681 cgaccgctgc aactcccggt gcaaccttgt cccggtctat tctcttcact gcaccagctc
     7741 caatctggtg tgaatgcccc tcgtctgttc gcgcaggcgg ggggctctat tcgtttgtca
     7801 gcatcgaaag tagccagatc agggatgcgt tgcaaccgcg tatgcccagg tcagaagagt
     7861 cgcacaagag ttgcagaccc ctggaaagaa aaatggccag agggcgaaaa caccctctga
     7921 ccagcggagc gggcgacggg aatcgaaccc gcgtagctag tttggaagaa tgggtgtctg
     7981 ccgaccacat atatgggccg gtcaagatag gtttttaccc cctctcggct gcatcctcta
     8041 agtggaaaga aattgcaggt cgtagaagcg cgttgaagcc tgagagttgc acaggagttg
     8101 caacccggta gccttgttca cgacgagagg agacctagtt ggcacgtcgc ggatggggat
     8161 cgctgaagac tcagcgcagc gggag
//
