LOCUS       sequence-12809-e        5300 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 5300)
  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 5300)
  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: pGMCKq1-0X1-sspBmyc
FEATURES             Location/Qualifiers
     source          1..5300
                     /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"
     CDS             complement(1187..2002)
                     /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     1910..1929
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1994..2013
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(3945..3991)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4248..4270
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4267..4283
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4267..4283
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4303..4323
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    4327..4351
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4740..4764)
                     /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(5293..5300,1..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 tagtgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca
      961 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     1021 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     1081 atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag
     1141 tcagcgtaat gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat
     1201 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1261 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1321 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     1381 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     1441 atggcaaaag attatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     1501 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     1561 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     1621 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     1681 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     1741 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     1801 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     1861 ggggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     1921 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg
     1981 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     2041 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     2101 caacgtcgct ttgttgctag ctggcgggaa cggcaatgct cctggcggcg gcggggctgg
     2161 tgcccaggtc tccgcacaga gcggcggtgc tggcgctcgc ggccaggcgt ggttcttcgc
     2221 gtactgacaa gaaacccccc tctttaggac tcagtgtcct tgggaggggg gctttttgcg
     2281 tttcaggagg tcttggccag cttggacatc gcctcagcga tagcctcgtc gcgggcctca
     2341 gacgccatct ggtacttcat cgccatccta ggagtcgtgt gaccgagacg ggccatcagc
     2401 tccttggtcg tcgcacctgc ctgagcggcg aacgtagcgc cgacagcgcg gaggtcgtgg
     2461 atgcggagtt ccggccgacc gatcttggcg tagccacgct tcagcgactt ggtgaacgcg
     2521 gacttcgaca gccggttgcc ctgcgtcgtg gtcaccagga atgcctcggg gcccttgttc
     2581 atcttcgtac ggtccttcat gtgcgctcgg atcatctccg cgacgtgagg cggaaccgtc
     2641 acaggacgct tcgaccggac ggtcttggcg ttgccaacga cgatcttgtt ccccacgcgg
     2701 gaagcgccac ggcgcacccg gagcttcatc gtcatgccgt cgtccacgat gtccttgcgg
     2761 cgaagctcga tcagctctcc gaaccggagg ctcgtccacg ccaggatgta tgccgcgatc
     2821 cggtagtgct cgaagatctc agcggcgacg atgtccagct cctcaggcgt cagcgcctct
     2881 acgtcgcgct catcggctgc cttctgctcg atccggcacg ggttctctgc gatcagcttg
     2941 tcctcgaccg ctgtgttcat caccgcccgg aggacgttgt aggcatgccg gcgggcagtc
     3001 gggtgcttcc tacccatccc ggcccaccac gcacgcacca gagctggcgt catctctgtg
     3061 accgccactt cacctagcac cgggtagatg cggcgctccg cgtgcccgct gtacagatcc
     3121 ctggtgccgt ctgcgaggtc gcgctccacg agccacttcc gggtgtactc ctccagcgtg
     3181 atggcgctgg cggctgcctt cttcgcccgg tcctgtggag gggtccaggt ctccatctcg
     3241 atgagccgct tctcgcccgc gagccaggct tcggcgtcca tcttgttgtc gtaggtctgc
     3301 agcgcgtagt acctcacacc gtcctgcggg ttgacgtatg aggcttggat cctcccgctg
     3361 cgctgagtct tcagcgatcc ccatccgcga cgtgccaact aggtctcctc tcgtcgtgaa
     3421 caaggctacc gggttgcaac tcctgtgcaa ctctcaggct tcaacgcgct tctacgacct
     3481 gcaatttctt tccacttaga ggatgcagcc gagagggggt aaaaacctat cttgaccggc
     3541 ccatatatgt ggtcggcaga cacccattct tccaaactag ctacgcgggt tcgattcccg
     3601 tcgcccgctc cgctggtcag agggtgtttt cgccctctgg ccatttttct ttccaggggt
     3661 ctgcaactct tgtgcgactc ttctgacctg ggcatacgcg gttgcaacgc atccctgatc
     3721 tggctacttt cgatgctgac aaacgaatag agccccccgc ctgcgcgaac agacgagggg
     3781 cattcacacc agattggagc tggtgcagtg aagagaatag accgggacaa ggttgcaccg
     3841 ggagttgcag cggtcggaac cctcgccgtc ggcgggctgg cgttcgccct gtcgttcacg
     3901 gctctagctg atcaccgcgg ccatgatggc cggacaaaca acagataaaa cgaaaggccc
     3961 agtctttcga ctgagccttt cgttttattt gatgcctggc agtcgatcgt acgctagtta
     4021 actacgtcga catcgattta attaatatgc atcaattgat ttaaatacta gtgttaacta
     4081 tttaattggg gaccctagag gtcccctttt ttattttaaa aattttttcn aaggggacct
     4141 ctagggtccc cagctggcta gcgagtcatg aggtnaccat gttctttcct gcgttatccc
     4201 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccagc ggataacaat
     4261 ttcacacagg aaacagctat gaccatgatt acgccaagct atcaactttg tatagaaaag
     4321 ttgaattcaa gtttgtacaa aaaagcaggc taccaggcct agatctgggg accctagagg
     4381 tccccttttt tattttaaaa attttttcac gtgccagccg attggactag aaatattgga
     4441 tcgtcggcac cgtcacggcc gtgggaggcg gcacgatccg cgacgtgatg atcggccgca
     4501 tccccacggt gctgcgcagt gagctctacg ccatcccggc gttgatctgt gcgttcgcac
     4561 gcacaggccc ggtgtgagaa gggtctctga cgagcgggag aactccctat cagtgataga
     4621 gtttgtcctc cctatcagtg atagataggc tctgggagta cccgtgtgta cgaccagcac
     4681 ggcatacatc atttcgacgc cgagagattc gccgcccgaa atgagcacga tccgcatgca
     4741 cccagctttc ttgtacaaag tgggctcgag gcatgcgaag gagatataca tatggacctg
     4801 tcgcagctga ccccgcgccg gccctacctg ctgcgcgcct tctacgagtg gttgctggac
     4861 aaccagctga ccccgcacct ggtggtggac gtgaccctgc cgggcgtgca ggtgccgatg
     4921 gagtacgcgc gcgacggcca gatcgtgctg aacatcgcgc cgcgcgccgt cggcaacctg
     4981 gagctggcga acgacgaggt gcgcttcaac gcgcgtttcg gtggcatccc gcggcaggtc
     5041 tcggtgccgc tggcggccgt gctggccatc tacgcccgcg agaacggcgc cggcaccatg
     5101 ttcgagccgg aggcggccta cgacgaggac acctcgatca tgaacgacga ggaggcctcg
     5161 gccgacaacg agaccgtgat gtcggtgatc gacggcgaca agccggacca cgacgatgac
     5221 acccacccgg acgatgagcc gccccagccg ccgcgcggcg gtcgcccggc cctgcgcgtc
     5281 gtgaagtaat agcaacttta
//
