LOCUS       sequence-12808-e        6690 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 6690)
  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 6690)
  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-luxAB-DAS+4
FEATURES             Location/Qualifiers
     source          1..6690
                     /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     3641..3663
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3660..3676
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3660..3676
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3696..3716
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3720..3744
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4236..4260)
                     /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(6683..6690,1..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 ttcaagggga cctctagggt
     3541 ccccagctgg ctagcgagtc atgaggtcac catgttcttt cctgcgttat cccctgattc
     3601 tgtggataac cgtattaccg cctttgagtg agctgatacc agcggataac aatttcacac
     3661 aggaaacagc tatgaccatg attacgccaa gctatcaact ttgtatagaa aagttgaatt
     3721 caagtttgta caaaaaagca ggcttattgg atcacgccgc gagcgtgtcg gtgttcgaag
     3781 acctgggaat ccagcaggcg cccaacagca ccgtctgctc cgtcttctcc agaaaggcga
     3841 ctacgcggtc ctcctcgtcg gtgggtacgc agccgaatgc ccgcgggtcg cccacccgca
     3901 ccagttgcag cgtgacatcg gctcgattgc ttcggtggaa gtccagcagt tgggccagat
     3961 ccgcgagggc cccccgtgaa catcgtcgtg gtaaccagcg tgtcggcttt ggctgtggcc
     4021 gtggtgcatt cggtggcgtt tgcgatcaag gggcgtcacg gcgaggacct ggtcgaccgc
     4081 tggaaccgtg agggcatcgg tgcacaccgc gggatcagac cgtcgacgaa ccatttgaat
     4141 ccgaccggta cctcaacgag ttgacggccg atcacgtcca ggttatgttc gatgacgatc
     4201 accgtattgc ccttgtcgac caggccgttg atccgaccca gctttcttgt acaaagtggt
     4261 taattaacag gatccccaaa taaggaaatg ttatgaaatt tggaaacttc cttctcactt
     4321 atcagccacc tgagctatct cagaccgaag tgatgaagcg attggttaat ctgggcaaag
     4381 cgtctgaagg ttgtggcttc gacaccgttt ggttgctaga gcaccacttc actgaatttg
     4441 ggttgttagg gaatccttat gttgctgccg cacacctatt aggtgcgaca gaaacgctca
     4501 acgttggcac tgcagctatc gtattgccga ctgcccatcc ggttcgacaa gcagaagacg
     4561 taaacctact ggatcaaatg tcaaaaggac gattccgttt tggtatttgt cgcggtttgt
     4621 acgataaaga ttttcgtgtc tttggtacag acatggataa cagccgagcc ttaatggact
     4681 gttggtatga cttgatgaaa gaaggcttca atgaaggcta tatcgcggcg gataacgaac
     4741 atattaagtt cccgaaaatc caactgaatc catcggctta cacacaaggt ggtgctcctg
     4801 tttatgtcgt cgcggagtca gcatcaacga cagaatgggc tgcagagcgt ggcctaccaa
     4861 tgattctaag ctggatcatc aacactcacg agaagaaagc gcagcttgat ctttacaacg
     4921 aagtcgcgac tgaacatggc tacgatgtga ctaagattga ccactgtttg tcttacatca
     4981 cctccgtcga tcatgactca aatagagcca aagatatttg ccgcaacttc ttgggccatt
     5041 ggtacgactc atacgtgaat gccaccaaga tttttgacga ctctgaccaa acaaaaggtt
     5101 acgacttcaa taaaggtcaa tggcgtgatt ttgtgttgaa aggccacaaa gacaccaatc
     5161 gccgaattga ttacagctac gaaatcaacc cagtagggac gcctgaagag tgtatcgcga
     5221 ttatccagca agatattgat gcgacgggta ttgacaatat ttgttgtggt tttgaagcaa
     5281 acggttctga agaagaaatt atcgcatcta tgaagctatt ccagtctgat gtgatgccat
     5341 atctcaaaga aaaacagtaa ttaatatttt ctaaaaggaa agagacatga aatttggatt
     5401 attcttcctc aattttatga attcaaagcg ttcttctgat caagtcatcg aagaaatgtt
     5461 agataccgca cattacgtag atcagttgaa gtttgacacg ttggctgttt acgaaaacca
     5521 tttctcgaac aatggtgtgg ttggtgcccc actaacagtg gctggttttt tacttggtat
     5581 gacaaagaac gccaaagtgg cttcgttgaa tcacgtcatt accacgcatc atccagtacg
     5641 tgtggcggaa gaagcgtgtc tacttgacca aatgagtgaa ggccgttttg cctttggctt
     5701 tagtgattgt gaaaagagtg cagatatgcg cttctttaat cgaccaacgg attctcagtt
     5761 tcagttgttc agtgagtgtc acaagatcat caatgatgca ttcactactg ggtactgcca
     5821 tccaaacaat gatttttata gttttcctaa aatctccgtt aacccacacg cgttcactga
     5881 aggcggtcct gcgcaatttg tgaatgcgac gagcaaagaa gtggttgaat gggcggctaa
     5941 gttagggctt ccactcgtgt ttagatggga cgactcaaac gctcaaagaa aagaatacgc
     6001 cggtttgtac cacgaagttg ctcaggcaca tggtgtcgat gttagtcagg ttcgacacaa
     6061 gctgacgctg ctggtcaacc aaaatgtaga tggtgaagca gcaagggcag aagctcgcgt
     6121 gtatttggaa gagtttgtcc gtgaatctta ctcaaatacc gactttgagc aaaaaatggg
     6181 agagctgttg tcagaaaatg ccatcggtac ttatgaagaa agtactcagg cagcgcgagt
     6241 tgcgattgag tgttgtggtg ccgcggacct attgatgtct tttgagtcga tggaagataa
     6301 agcgcagcaa agagcggtta tcgatgtggt aaacgccaac atcgtcaaat accactcggc
     6361 cgccaacgac gagaactact ccgagaacta cgcggacgcc agctaataac tagtcgttta
     6421 actgatgctg aaggggcagc gatgcccctt atatcaccat tcttttcgcg cgatagcgct
     6481 aactaataga ggcatttata tggacgtact ttcagcggtt aagcaggaaa atatcgcagc
     6541 gagcacagaa atcgatgact tgattttcat gggaactcct cagcaatggt cattgcagga
     6601 acaaaaaaca gccggatctc gagctcgcga aagctctgat ttaaatggat cccccgggct
     6661 gcaggaattc gatatcaagc ttcaacttta
//
