LOCUS       sequence-12918-e        7937 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 7937)
  AUTHORS   Klotzsche M, Ehrt S, Schnappinger D
  TITLE     Improved tetracycline repressors for gene silencing in mycobacteria.
  JOURNAL   Nucleic Acids Res. 2009 Apr;37(6):1778-88. doi: 10.1093/nar/gkp015. 
            Epub 2009 Jan 27.
  PUBMED    19174563
REFERENCE   2  (bases 1 to 7937)
  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: 
            "10.1093/nar/gkp015"; journalName: "Nucleic Acids Res"; date: 
            "2009-04"; volume: "37"; issue: "6"; pages: "1778-88"
COMMENT     Sequence Label: pGMCK-P38-luxAB-DAS+4
FEATURES             Location/Qualifiers
     source          1..7937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(21..37)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(30..52)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    596..730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(633..652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     687..706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(926..1514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1006..1025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1846..2661)
                     /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"
     primer_bind     2569..2588
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2653..2672
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(4604..4650)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4907..4929
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4926..4942
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4926..4942
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4962..4982
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    4986..5010
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5502..5526)
                     /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(7930..7937,1..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 ttgataattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
       61 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      121 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggggattg
      241 gtggcgacga ctcctggagc ccgtcagtat cggcggaatt ccagctgagc gccggtcgct
      301 accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca tgtctgcccg
      361 tatttcgcgt aaggaaatcc attatgtact atttaaaaaa cacaaacttt tggatgttcg
      421 gtttattctt tttcttttac ttttttatca tgggagccta cttcccgttt ttcccgattt
      481 ggctacatga catcaaccat atcagcaaaa gtgatacggg tattattttt gccgctattt
      541 ctctgttctc gctattattc caaccgctgt ttggtctgct ttctgacaaa ctcggaactt
      601 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      661 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      721 tgtctggatc ctctagagtc gacctgcagg tttaaacgcg gccgcacgcg ttgcgctcgg
      781 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      841 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      901 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      961 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1021 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1081 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     1141 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1201 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1261 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1321 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1381 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1441 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1501 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaact
     1561 agtgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat
     1621 catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt
     1681 tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga
     1741 tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt
     1801 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     1861 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     1921 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     1981 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2041 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2101 tggcaaaaga ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2161 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2221 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     2281 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     2341 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     2401 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     2461 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatg
     2521 gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     2581 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt
     2641 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     2701 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     2761 aacgtcgctt tgttgctagc tggcgggaac ggcaatgctc ctggcggcgg cggggctggt
     2821 gcccaggtct ccgcacagag cggcggtgct ggcgctcgcg gccaggcgtg gttcttcgcg
     2881 tactgacaag aaacccccct ctttaggact cagtgtcctt gggagggggg ctttttgcgt
     2941 ttcaggaggt cttggccagc ttggacatcg cctcagcgat agcctcgtcg cgggcctcag
     3001 acgccatctg gtacttcatc gccatcctag gagtcgtgtg accgagacgg gccatcagct
     3061 ccttggtcgt cgcacctgcc tgagcggcga acgtagcgcc gacagcgcgg aggtcgtgga
     3121 tgcggagttc cggccgaccg atcttggcgt agccacgctt cagcgacttg gtgaacgcgg
     3181 acttcgacag ccggttgccc tgcgtcgtgg tcaccaggaa tgcctcgggg cccttgttca
     3241 tcttcgtacg gtccttcatg tgcgctcgga tcatctccgc gacgtgaggc ggaaccgtca
     3301 caggacgctt cgaccggacg gtcttggcgt tgccaacgac gatcttgttc cccacgcggg
     3361 aagcgccacg gcgcacccgg agcttcatcg tcatgccgtc gtccacgatg tccttgcggc
     3421 gaagctcgat cagctctccg aaccggaggc tcgtccacgc caggatgtat gccgcgatcc
     3481 ggtagtgctc gaagatctca gcggcgacga tgtccagctc ctcaggcgtc agcgcctcta
     3541 cgtcgcgctc atcggctgcc ttctgctcga tccggcacgg gttctctgcg atcagcttgt
     3601 cctcgaccgc tgtgttcatc accgcccgga ggacgttgta ggcatgccgg cgggcagtcg
     3661 ggtgcttcct acccatcccg gcccaccacg cacgcaccag agctggcgtc atctctgtga
     3721 ccgccacttc acctagcacc gggtagatgc ggcgctccgc gtgcccgctg tacagatccc
     3781 tggtgccgtc tgcgaggtcg cgctccacga gccacttccg ggtgtactcc tccagcgtga
     3841 tggcgctggc ggctgccttc ttcgcccggt cctgtggagg ggtccaggtc tccatctcga
     3901 tgagccgctt ctcgcccgcg agccaggctt cggcgtccat cttgttgtcg taggtctgca
     3961 gcgcgtagta cctcacaccg tcctgcgggt tgacgtatga ggcttggatc ctcccgctgc
     4021 gctgagtctt cagcgatccc catccgcgac gtgccaacta ggtctcctct cgtcgtgaac
     4081 aaggctaccg ggttgcaact cctgtgcaac tctcaggctt caacgcgctt ctacgacctg
     4141 caatttcttt ccacttagag gatgcagccg agagggggta aaaacctatc ttgaccggcc
     4201 catatatgtg gtcggcagac acccattctt ccaaactagc tacgcgggtt cgattcccgt
     4261 cgcccgctcc gctggtcaga gggtgttttc gccctctggc catttttctt tccaggggtc
     4321 tgcaactctt gtgcgactct tctgacctgg gcatacgcgg ttgcaacgca tccctgatct
     4381 ggctactttc gatgctgaca aacgaataga gccccccgcc tgcgcgaaca gacgaggggc
     4441 attcacacca gattggagct ggtgcagtga agagaataga ccgggacaag gttgcaccgg
     4501 gagttgcagc ggtcggaacc ctcgccgtcg gcgggctggc gttcgccctg tcgttcacgg
     4561 ctctagctga tcaccgcggc catgatggcc ggacaaacaa cagataaaac gaaaggccca
     4621 gtctttcgac tgagcctttc gttttatttg atgcctggca gtcgatcgta cgctagttaa
     4681 ctacgtcgac atcgatttaa ttaatatgca tcaattgatt taaatactag tgttaactat
     4741 ttaattgggg accctagagg tccccttttt tattttaaaa attttttcna aggggacctc
     4801 tagggtcccc agctggctag cgagtcatga ggtnaccatg ttctttcctg cgttatcccc
     4861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccagcg gataacaatt
     4921 tcacacagga aacagctatg accatgatta cgccaagcta tcaactttgt atagaaaagt
     4981 tgaattcaag tttgtacaaa aaagcaggct tattggatca cgccgcgagc gtgtcggtgt
     5041 tcgaagacct gggaatccag caggcgccca acagcaccgt ctgctccgtc ttctccagaa
     5101 aggcgactac gcggtcctcc tcgtcggtgg gtacgcagcc gaatgcccgc gggtcgccca
     5161 cccgcaccag ttgcagcgtg acatcggctc gattgcttcg gtggaagtcc agcagttggg
     5221 ccagatccgc gagggccccc cgtgaacatc gtcgtggtaa ccagcgtgtc ggctttggct
     5281 gtggccgtgg tgcattcggt ggcgtttgcg atcaaggggc gtcacggcga ggacctggtc
     5341 gaccgctgga accgtgaggg catcggtgca caccgcggga tcagaccgtc gacgaaccat
     5401 ttgaatccga ccggtacctc aacgagttga cggccgatca cgtccaggtt atgttcgatg
     5461 acgatcaccg tattgccctt gtcgaccagg ccgttgatcc gacccagctt tcttgtacaa
     5521 agtggttaat taacaggatc ccaaataagg aaatgttatg aaatttggaa acttccttct
     5581 cacttatcag ccacctgagc tatctcagac cgaagtgatg aagcgattgg ttaatctggg
     5641 caaagcgtct gaaggttgtg gcttcgacac cgtttggttg ctagagcacc acttcactga
     5701 atttgggttg ttagggaatc cttatgttgc tgccgcacac ctattaggtg cgacagaaac
     5761 gctcaacgtt ggcactgcag ctatcgtatt gccgactgcc catccggttc gacaagcaga
     5821 agacgtaaac ctactggatc aaatgtcaaa aggacgattc cgttttggta tttgtcgcgg
     5881 tttgtacgat aaagattttc gtgtctttgg tacagacatg gataacagcc gagccttaat
     5941 ggactgttgg tatgacttga tgaaagaagg cttcaatgaa ggctatatcg cggcggataa
     6001 cgaacatatt aagttcccga aaatccaact gaatccatcg gcttacacac aaggtggtgc
     6061 tcctgtttat gtcgtcgcgg agtcagcatc aacgacagaa tgggctgcag agcgtggcct
     6121 accaatgatt ctaagctgga tcatcaacac tcacgagaag aaagcgcagc ttgatcttta
     6181 caacgaagtc gcgactgaac atggctacga tgtgactaag attgaccact gtttgtctta
     6241 catcacctcc gtcgatcatg actcaaatag agccaaagat atttgccgca acttcttggg
     6301 ccattggtac gactcatacg tgaatgccac caagattttt gacgactctg accaaacaaa
     6361 aggttacgac ttcaataaag gtcaatggcg tgattttgtg ttgaaaggcc acaaagacac
     6421 caatcgccga attgattaca gctacgaaat caacccagta gggacgcctg aagagtgtat
     6481 cgcgattatc cagcaagata ttgatgcgac gggtattgac aatatttgtt gtggttttga
     6541 agcaaacggt tctgaagaag aaattatcgc atctatgaag ctattccagt ctgatgtgat
     6601 gccatatctc aaagaaaaac agtaattaat attttctaaa aggaaagaga catgaaattt
     6661 ggattattct tcctcaattt tatgaattca aagcgttctt ctgatcaagt catcgaagaa
     6721 atgttagata ccgcacatta cgtagatcag ttgaagtttg acacgttggc tgtttacgaa
     6781 aaccatttct cgaacaatgg tgtggttggt gccccactaa cagtggctgg ttttttactt
     6841 ggtatgacaa agaacgccaa agtggcttcg ttgaatcacg tcattaccac gcatcatcca
     6901 gtacgtgtgg cggaagaagc gtgtctactt gaccaaatga gtgaaggccg ttttgccttt
     6961 ggctttagtg attgtgaaaa gagtgcagat atgcgcttct ttaatcgacc aacggattct
     7021 cagtttcagt tgttcagtga gtgtcacaag atcatcaatg atgcattcac tactgggtac
     7081 tgccatccaa acaatgattt ttatagtttt cctaaaatct ccgttaaccc acacgcgttc
     7141 actgaaggcg gtcctgcgca atttgtgaat gcgacgagca aagaagtggt tgaatgggcg
     7201 gctaagttag ggcttccact cgtgtttaga tgggacgact caaacgctca aagaaaagaa
     7261 tacgccggtt tgtaccacga agttgctcag gcacatggtg tcgatgttag tcaggttcga
     7321 cacaagctga cgctgctggt caaccaaaat gtagatggtg aagcagcaag ggcagaagct
     7381 cgcgtgtatt tggaagagtt tgtccgtgaa tcttactcaa ataccgactt tgagcaaaaa
     7441 atgggagagc tgttgtcaga aaatgccatc ggtacttatg aagaaagtac tcaggcagcg
     7501 cgagttgcga ttgagtgttg tggtgccgcg gacctattga tgtcttttga gtcgatggaa
     7561 gataaagcgc agcaaagagc ggttatcgat gtggtaaacg ccaacatcgt caaataccac
     7621 tcggccgcca acgacgagaa ctactccgag aactacgcgg acgccagcta ataactagtc
     7681 gtttaactga tgctgaaggg gcagcgatgc cccttatatc accattcttt tcgccgatag
     7741 cgctaactaa tagaggcatt tatatggacg tactttcagc ggttaagcag gaaaacatcg
     7801 cagcgagcac agaaatcgat gacttgattt tcatgggaac tcctcagcaa tggtcattgc
     7861 aggaacaaaa acagctgaag ctctgattta aatggatccc ccgggctgca ggaattcgat
     7921 atcaagcttc aacttta
//
