LOCUS       sequence-73901-e        6218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nourseothricin resistant; expresses sspB under the control of a 
            promoter containing PtetO and constitutively expressses single-chain
            TetR10 (tsc10); integrates into mycobacteriophage att-Tweety site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Kim JH, O'Brien KM, Sharma R, Boshoff HI, Rehren G, Chakraborty S, 
            Wallach JB, Monteleone M, Wilson DJ, Aldrich CC, Barry CE 3rd, Rhee 
            KY, Ehrt S, Schnappinger D
  TITLE     A genetic strategy to identify targets for the development of drugs 
            that prevent bacterial persistence.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Nov 4.
  PUBMED    24191058
REFERENCE   2  (bases 1 to 6218)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Nov 4."
COMMENT     Sequence Label: pGMCtN-TSC10M1-sspB
FEATURES             Location/Qualifiers
     source          1..6218
                     /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    5245..5269
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5658..5682)
                     /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(6211..6218,1..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 aaagttgcag
     3721 ctggctagcg agtcatgagg tcaccatggt cattaggagc cgctctcgca cttcagctgc
     3781 ttctccaagc cgcagatgat cagctccagg ccgaacaaga aggccggctc ggcgccctgg
     3841 tggtcgaaca gctcgatggc ctggcgcagc aacggcggca tgctgtcggt ggtcggggtc
     3901 tcgcgctcct ccttggcgac ctggtgctcc tggtcctcca acacgcagcc cagggtgaag
     3961 tggcccacgg ccgacagggc gtacaacgcg ttctccagcg agaagccctg ctggcacagg
     4021 aaggccaact ggttctccag ggtctcgtac tgcttctcgg tcggccgggt gcccaggtgc
     4081 accttggcgc cgtcgcggtg gctcagcaag gcgcagcgga agctcttggc gttgttgcgc
     4141 aggaagtcct gccagctctc gccctccagc gggcagaagt gggtgtggtg gcggtccaac
     4201 atctcgatgg ccagggcgtc gagcagggcc cgcttgttct tgacgtgcca gtacagggtc
     4261 ggctgctcga cgcccagctt ctgggccagc ttgcgggtgg tcaggccctc gatgccgacc
     4321 tcgttcagca gctccagggc ggagttgatg accttggact tgtccaggcg ggatctagag
     4381 ccacctccgc ccgatccgcc acctccagag cctccaccgc cagaaccgcc tccaccggat
     4441 ccaccgcctc ctgagtcgcc cgactcacac ttcaactgct tctccagccc acaaatgatg
     4501 agttccaaac cgaacaggaa cgcgggctcc gcaccctgat gatcgaagag ctcgatcgcc
     4561 tgccggagca gcgggggcat cgaatccgtg gtgggcgtct cccgctcttc cttcgccacc
     4621 tgatgttctt ggtcttcgag gacgcacccc agcgtgaaat gcccgaccgc gctcagcgca
     4681 tacagggcgt tctcgaggga gaacccttgc tgacacaaga acgcgagctg gttctcgagc
     4741 gtttcatact gcttctccgt gggccgcgta ccgagatgga ccttcgcccc gtcccgatgg
     4801 gagaggagcg cacaccggaa ggacttcgcg ttgttccgga ggaagtcttg ccacgactcc
     4861 ccttccaagg gacagaagtg cgtatgatgc cggtcgagca tttcgatcgc gagcgcatcc
     4921 agcaacgcgc gcttgttctt cacatgccaa tacaacgtgg gttgctccac gcccaacttc
     4981 tgcgcgagct tccgggtcgt caacccttcg atgcccacct cattcaacag ttccaacgcg
     5041 ctgttgatca ccttgctctt atccaaccgc gacattgcga agtgattcct cctgatggta
     5101 ctggtaccgg atccgtggca ggagggtcgc ctgcgtcggc aactcgccca ggctacacga
     5161 gagccctcga ctgcgaaaat gccgaccagg caccaccgca ctccaggctt aacccggtgc
     5221 gacgatccaa tattacgaga tctgacaagt ttgtacaaaa aagcaggcta ccaggcctag
     5281 atctggggac cctagaggtc ccctttttta ttttaaaaat tttttcacgt gccagccgat
     5341 tggactagaa atattggatc gtcggcaccg tcacggccgt gggaggcggc acgatccgcg
     5401 acgtgatgat cggccgcatc cccacggtgc tgcgcagtga gctctacgcc atcccggcgt
     5461 tgatctgtgc gttcgcacgc acaggcccgg tgtgagaagg gtctctgacg agcgggagaa
     5521 ctccctatca gtgatagagt ttgtcctccc tatcagtgat agataggctc tgggagtacc
     5581 cgtgtgtacg accagcacgg catacatcat ttcgacgccg agagattcgc cgcccgaaat
     5641 gagcacgatc cgcatgcacc cagctttctt gtacaaagtg ggctcgaggc atgcgaagga
     5701 gatatacata tggacctgtc gcagctgacc ccgcgccggc cctacctgct gcgcgccttc
     5761 tacgagtggt tgctggacaa ccagctgacc ccgcacctgg tggtggacgt gaccctgccg
     5821 ggcgtgcagg tgccgatgga gtacgcgcgc gacggccaga tcgtgctgaa catcgcgccg
     5881 cgcgccgtcg gcaacctgga gctggcgaac gacgaggtgc gcttcaacgc gcgtttcggt
     5941 ggcatcccgc ggcaggtctc ggtgccgctg gcggccgtgc tggccatcta cgcccgcgag
     6001 aacggcgccg gcaccatgtt cgagccggag gcggcctacg acgaggacac ctcgatcatg
     6061 aacgacgagg aggcctcggc cgacaacgag accgtgatgt cggtgatcga cggcgacaag
     6121 ccggaccacg acgatgacac ccacccggac gatgagccgc cccagccgcc gcgcggcggt
     6181 cgcccggccc tgcgcgtcgt gaagtaatag caacttta
//
