LOCUS       sequence-12806-e        6620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  AUTHORS   Kim JH, Wei JR, Wallach JB, Robbins RS, Rubin EJ, Schnappinger D
  TITLE     Protein inactivation in mycobacteria by controlled proteolysis and 
            its application to deplete the beta subunit of RNA polymerase.
  JOURNAL   Nucleic Acids Res. 2011 Mar;39(6):2210-20. doi: 10.1093/nar/gkq1149
  PUBMED    21075796
REFERENCE   2  (bases 1 to 6620)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2011-03"; volume: "39"; issue: "6"; 
            pages: "2210-20"
COMMENT     Sequence Label: pGMCtN-P38-luxAB2
FEATURES             Location/Qualifiers
     source          1..6620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(267..855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(347..366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(932..1018)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1063..1632)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     primer_bind     3642..3664
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3661..3677
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3661..3677
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3697..3717
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3721..3745
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4237..4261)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(6613..6620,1..13))
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 ttatacatag ttgcttaagc tgcaggggac cctagaggtc ccctttttta ttttaaaaat
       61 tttttataac cggtaagaag tccaacacgc cgctcctccc cgatgttgcg tagccgcttc
      121 aagcttaagc aacggcccca gtgagtagcg actttggatc cggtacccgc aacggtcaac
      181 cagcgtctac gcgtttcact ggccgacatg tgagcaaaag gccagcaaaa ggccaggaac
      241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
      481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
      661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
      721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
      781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      901 tagtcgcaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
      961 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     1021 gcctggcagt tccctactct cgcatgggga gagatgcatc tatcaggggc agggcatgct
     1081 catgtagagc gcctgctcgc cgtccgaggc ggtgccgtcg tacagggcgg tgtccaggcc
     1141 gcagagggtg aaccccatcc gccggtacgc gtggatcgcc ggtgcgttga cgttggtgac
     1201 ctccagccag aggtgcccgg cgccccgctc gcgggcgaac tccgtcgcga gccccatcaa
     1261 cgcgcgcccg accccgtgcc cccggtgctc cggggcgacc tcgatgtcct cgacggtcag
     1321 ccggcggttc cagccggagt acgagacgac cacgaagccc gccaggtcgc cgtcgtcccc
     1381 gtacgcgacg aacgtccggg agtccgggtc gccgtcctcc ccggcgtccg attcgtcgtc
     1441 cgattcgtcg tcggggaaca ccttggtcag gggcgggtcc accggcacct cccgcagggt
     1501 gaagccgtcc ccggtggcgg tgacgcggaa gacggtgtcg gtggtgaagg acccatccag
     1561 tgcctcgatg gcctcggcgt cccccgggac actggtgcgg taccggtaag ccgtgtcgtc
     1621 aagagtggtc atatgtatat ctccttcgca tgcctcccag attatatcac tcgagctagc
     1681 tggtgtcaag gcgcgcctgc aggtcgagat ttacagtcgg tcgcctacgc aaacgtcgat
     1741 gggagcagtt cggacagccc ctgcatggcc tccagatgcc tcgcccgatc cgcatgcgca
     1801 tagatccgct gcgcatccac actcgcatga cccaagatct ccatacgcgt ttgctcatcc
     1861 acacccgctg cgcgcagcaa tgtcgaggtg gtgtgccgcg agttgtgcgg cggtaacgac
     1921 tcggttggac cgatcacccc agcagcgcgg aacacgccac gccacacgtc gtagtccgaa
     1981 cggggatcga tcggcttccc ctccttgtgc cacaccaagt catgcggatt gtcggtgcgg
     2041 agtttctgca tcgccacata caacggcggc aacaacggaa cctcacgcca accagcgtcc
     2101 gtcttcggcc gggtgaacaa caacgacccc tcacattcct gatactcgaa atgcgccggc
     2161 aggtcccacc gggactgcgg gcatgcccat gcccgtgtct tcccgcaagg ccagtacggg
     2221 ggtttcttgg gcatacggtc gggccgggac agcggtgacg gttcgggtag aggatcccca
     2281 cagccgtgga cgcgggtttc cgattgcaac tgccaagcga tggtgatcca tccctgagcg
     2341 gggttgtcga cgtagggcca gcgcaggccg aggagttccc cacggcgggc gcccgtcagg
     2401 aaaccggcgg cgatccgcac cgcatccggt tcgtcgcaca cctggaacgc ggtgtgaatg
     2461 atgtgctgcg ccacgtccgc cgggaagccg ttgcgtttct tcttccggta ctcgggcttg
     2521 tcgaccaatg cggccacatt cctggtcgcc acaccctccg ctaccgcatc gtccaaggct
     2581 ttctggacga tgacatggac cagctcggcg gtgcgggagg ccccgatctc ggagtgcagg
     2641 tcccgcacat gctgcggggt gagtttgtcg atgcgtttcg cgccgaggat cgggttgatg
     2701 tggttgtgga tggcggcccg gtagtcgttg aggacgccgg ggcggacttt acgtttggcg
     2761 tggatgttgt cgatccagtg cagcatccac ttctccacag ttgtggatga ggtggtggcg
     2821 atgcggccct cttcgacgtc gcggcggagt tgtttgagtt tggccatggc ggtgttgcgg
     2881 tcaacggagg acacccattt gtagcggcgg ttgccgttgc ggtctggggg tagttctact
     2941 cgtcccatcc atttgccgtc ggcgcgttgg aagaacgctc cgtctccgcg ggttctgcgt
     3001 ttcttagttg ccatcgtttt ccctcccagg gggtcaccct acggttcacc ctacggtgct
     3061 gcgcagcatt acgcagaatt gcgcagtatc gggggtctac ctgcgggttt gacaacgttt
     3121 ctcctggtat gcagcctatc aaccgctgac tcttaatcag cgggtcgggg gttcgaaacc
     3181 ctcacggcgc acaggtcaga ggccataagc ctcagagggg atcaccctaa aggtaaccct
     3241 ataggggttt tcactggtcc tacatgtcgt cgcgccgtcg tactctcttt tcatagggga
     3301 gcatgaggat ccaactgagc ttgctgtgcc ttcgattgag cggttgaagc atctggagcg
     3361 tcggcggaca gcgatcacgg ttcccaacga tggctgaaag aaggatgaaa tgactgcact
     3421 catcgataag cttcgaattc tgcagctgga tccatggata tctagtgtta actatttaat
     3481 tggggaccct agaggtcccc ttttttattt taaaaatttt ttcnaagggg acctctaggg
     3541 tccccagctg gctagcgagt catgaggtna ccatgttctt tcctgcgtta tcccctgatt
     3601 ctgtggataa ccgtattacc gcctttgagt gagctgatac cagcggataa caatttcaca
     3661 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaat
     3721 tcaagtttgt acaaaaaagc aggcttattg gatcacgccg cgagcgtgtc ggtgttcgaa
     3781 gacctgggaa tccagcaggc gcccaacagc accgtctgct ccgtcttctc cagaaaggcg
     3841 actacgcggt cctcctcgtc ggtgggtacg cagccgaatg cccgcgggtc gcccacccgc
     3901 accagttgca gcgtgacatc ggctcgattg cttcggtgga agtccagcag ttgggccaga
     3961 tccgcgaggg ccccccgtga acatcgtcgt ggtaaccagc gtgtcggctt tggctgtggc
     4021 cgtggtgcat tcggtggcgt ttgcgatcaa ggggcgtcac ggcgaggacc tggtcgaccg
     4081 ctggaaccgt gagggcatcg gtgcacaccg cgggatcaga ccgtcgacga accatttgaa
     4141 tccgaccggt acctcaacga gttgacggcc gatcacgtcc aggttatgtt cgatgacgat
     4201 caccgtattg cccttgtcga ccaggccgtt gatccgaccc agctttcttg tacaaagtgg
     4261 ttaattaaca ggatccccaa ataaggaaat gttatgaaat ttggaaactt ccttctcact
     4321 tatcagccac ctgagctatc tcagaccgaa gtgatgaagc gattggttaa tctgggcaaa
     4381 gcgtctgaag gttgtggctt cgacaccgtt tggttgctag agcaccactt cactgaattt
     4441 gggttgttag ggaatcctta tgttgctgcc gcacacctat taggtgcgac agaaacgctc
     4501 aacgttggca ctgcagctat cgtattgccg actgcccatc cggttcgaca agcagaagac
     4561 gtaaacctac tggatcaaat gtcaaaagga cgattccgtt ttggtatttg tcgcggtttg
     4621 tacgataaag attttcgtgt ctttggtaca gacatggata acagccgagc cttaatggac
     4681 tgttggtatg acttgatgaa agaaggcttc aatgaaggct atatcgcggc ggataacgaa
     4741 catattaagt tcccgaaaat ccaactgaat ccatcggctt acacacaagg tggtgctcct
     4801 gtttatgtcg tcgcggagtc agcatcaacg acagaatggg ctgcagagcg tggcctacca
     4861 atgattctaa gctggatcat caacactcac gagaagaaag cgcagcttga tctttacaac
     4921 gaagtcgcga ctgaacatgg ctacgatgtg actaagattg accactgttt gtcttacatc
     4981 acctccgtcg atcatgactc aaatagagcc aaagatattt gccgcaactt cttgggccat
     5041 tggtacgact catacgtgaa tgccaccaag atttttgacg actctgacca aacaaaaggt
     5101 tacgacttca ataaaggtca atggcgtgat tttgtgttga aaggccacaa agacaccaat
     5161 cgccgaattg attacagcta cgaaatcaac ccagtaggga cgcctgaaga gtgtatcgcg
     5221 attatccagc aagatattga tgcgacgggt attgacaata tttgttgtgg ttttgaagca
     5281 aacggttctg aagaagaaat tatcgcatct atgaagctat tccagtctga tgtgatgcca
     5341 tatctcaaag aaaaacagta attaatattt tctaaaagga aagagacatg aaatttggat
     5401 tattcttcct caattttatg aattcaaagc gttcttctga tcaagtcatc gaagaaatgt
     5461 tagataccgc acattacgta gatcagttga agtttgacac gttggctgtt tacgaaaacc
     5521 atttctcgaa caatggtgtg gttggtgccc cactaacagt ggctggtttt ttacttggta
     5581 tgacaaagaa cgccaaagtg gcttcgttga atcacgtcat taccacgcat catccagtac
     5641 gtgtggcgga agaagcgtgt ctacttgacc aaatgagtga aggccgtttt gcctttggct
     5701 ttagtgattg tgaaaagagt gcagatatgc gcttctttaa tcgaccaacg gattctcagt
     5761 ttcagttgtt cagtgagtgt cacaagatca tcaatgatgc attcactact gggtactgcc
     5821 atccaaacaa tgatttttat agttttccta aaatctccgt taacccacac gcgttcactg
     5881 aaggcggtcc tgcgcaattt gtgaatgcga cgagcaaaga agtggttgaa tgggcggcta
     5941 agttagggct tccactcgtg tttagatggg acgactcaaa cgctcaaaga aaagaatacg
     6001 ccggtttgta ccacgaagtt gctcaggcac atggtgtcga tgttagtcag gttcgacaca
     6061 agctgacgct gctggtcaac caaaatgtag atggtgaagc agcaagggca gaagctcgcg
     6121 tgtatttgga agagtttgtc cgtgaatctt actcaaatac cgactttgag caaaaaatgg
     6181 gagagctgtt gtcagaaaat gccatcggta cttatgaaga aagtactcag gcagcgcgag
     6241 ttgcgattga gtgttgtggt gccgcggacc tattgatgtc ttttgagtcg atggaagata
     6301 aagcgcagca aagagcggtt atcgatgtgg taaacgccaa catcgtcaaa taccactcgt
     6361 aacgtttaac tgatgctgaa ggggcagcga tgccccttat atcaccattc ttttcgccga
     6421 tagcgctaac taatagaggc atttatatgg acgtactttc agcggttaag caggaaaaca
     6481 tcgcagcgag cacagaaatc gatgacttga ttttcatggg aactcctcag caatggtcat
     6541 tgcaggaaca aaaacagctg aagctctgat ttaaatggat cccccgggct gcaggaattc
     6601 gatatcaagc ttcaacttta
//
