LOCUS       sequence-211179-        8906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pMINTC3GH but containing oriM for episomal replication 
            in mycobacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8906)
  AUTHORS   Arnold FM, Hohl M, Remm S, Koliwer-Brandl H, Adenau S, Chusri S, 
            Sander P, Hilbi H, Seeger MA
  TITLE     A uniform cloning platform for mycobacterial genetics and protein 
            production.
  JOURNAL   Sci Rep. 2018 Jun 22;8(1):9539. doi: 10.1038/s41598-018-27687-5.
  PUBMED    29934571
REFERENCE   2  (bases 1 to 8906)
  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: "Sci Rep."; 
            date: "2018-06-22"; volume: "8(1):9539. doi"; pages: " 
            10.1038/s41598-018-27687-5"
COMMENT     Sequence Label: pMEXC3GH (Apr)
FEATURES             Location/Qualifiers
     source          1..8906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(300..324)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(403..708)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(505..524)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1050..1709)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1624..1643
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1763..1769,1770..1787,1788..1793))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1818..1942)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1961..1981)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1985..2005)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2032..2055
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2062..2772
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2089..2117)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2712..2732
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2779..2808
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     rep_origin      complement(3078..3666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3158..3177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4158..4958
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     primer_bind     5151..5170
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(7409..7455)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7711..7733
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7730..7746
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7730..7746
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    7766..7786
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7839..7855
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7840..7856
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8793..8817
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tcgtcggcac cgtcacggcc gtgggaggcg gcacgatccg cgacgtgatg atcggccgca
       61 tccccacggt gctgcgcagt gagctctacg ccatcccggc gttgatctgt gcgttcgcac
      121 gcacaggccc ggtgtgagaa gggtctctga cgagcgggag aactccctat cagtgataga
      181 gtttgtcctc cctatcagtg atagataggc tctgggagta cccgtgtgta cgaccagcac
      241 ggcatacatc atttcgacgc cgagagattc gccgcccgaa atgagcacga tccgcatgca
      301 cccagctttc ttgtacaaag tgggctcgag gcatgcgaag gagatataca tatgagttga
      361 agagcgacct gcagactggc tgtgtataag ggagcctgac atttatattc cccagaacat
      421 caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc
      481 gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc
      541 gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc
      601 cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta catccacaaa
      661 cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt caccagcccc
      721 tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca gccatccctt
      781 cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc tgcatggttg
      841 tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat agctgtcgct
      901 gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat acttcgggta
      961 tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc atactgttat
     1021 ctggctttta gtaagccgga tccacgcgtt tacgccccgc cctgccactc atcgcagtac
     1081 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     1141 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     1201 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     1261 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     1321 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     1381 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     1441 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     1501 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     1561 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     1621 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     1681 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgac
     1741 ggatcctaac tcaaaatcca cacattatac gagccggaag cataaagtgt aaagcctggg
     1801 gtgcctaatg cggccgccat agtgactgga tatgttgtgt tttacagtat tatgtagtct
     1861 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt
     1921 cagctttttt gtacaaactt gtgatatcga attcgctagc tacctttctc ttcttttttg
     1981 gatctacctt tctcttcttt tttggggatc cactagttca gctcttctgc attagaagtt
     2041 ttgtttcaag gtccacaatt cagcaaagga gaagaacttt tcactggagt tgtcccaatt
     2101 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     2161 ggtgatgcta catacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
     2221 gttccttggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
     2281 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     2341 gaacgcacta tatctttcaa agatgacggg aactacaaga cgcgtgctga agtcaagttt
     2401 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
     2461 aacattctcg gacacaaact agagtacaac tataactcac acaatgtata catcacggca
     2521 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacat tgaagatggt
     2581 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     2641 ttaccagaca accattacct gtcgacacaa tctgcccttt cgaaagatcc caacgaaaag
     2701 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
     2761 gagctctaca aaactagtca tcatcatcac catcatcatc accatcatta attatacata
     2821 gttgcttaag ctgcagggga ccctagaggt cccctttttt attttaaaaa ttttttataa
     2881 ccggtaagaa gtccaacacg ccgctcctcc ccgatgttgc gtagccgctt caagcttaag
     2941 caacggcccc agtgagtagc gactttggat ccggtacccg caacggtcaa ccagcgtcta
     3001 cgcgtttcac tggccgacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3241 tttctcccct cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3481 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcatggaac tagtgaggtc
     3721 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     3781 agaaagtgag ggagccacgg ttggtgagag ctttgttgta ggtggaccag ttggtgattt
     3841 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     3901 tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat
     3961 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     4021 atgaaactgc aagtatctag aaggatcttc acctagatcc ttttggttca tgtgcagctc
     4081 catcagcaaa aggggatgat aagtttatca ccaccgacta tttgcaacag tgccgttgat
     4141 cgtgctatga tcgactgatg tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc
     4201 gaaaagccga gctcatcggt cagcttctca accttggggt tacccccggc ggtgtgctgc
     4261 tggtccacag ctccttccgt agcgtccggc ccctcgaaga tgggccactt ggactgatcg
     4321 aggccctgcg tgctgcgctg ggtccgggag ggacgctcgt catgccctcg tggtcaggtc
     4381 tggacgacga gccgttcgat cctgccacgt cgcccgttac accggacctt ggagttgtct
     4441 ctgacacatt ctggcgcctg ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg
     4501 cagcggggcc acaggcagag cagatcatct ctgatccatt gcccctgcca cctcactcgc
     4561 ctgcaagccc ggtcgcccgt gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg
     4621 gacacgatgc caacacgacg ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg
     4681 tgccgagaca ctgcaccatt cttcaggatg gcaagttggt acgcgtcgat tatctcgaga
     4741 atgaccactg ctgtgagcgc tttgccttgg cggacaggtg gctcaaggaa aagagccttc
     4801 agaaggaagg tccagtcggt catgcctttg ctcggttgat ccgctcccgc gacattgtgg
     4861 cgacagccct gggtcaactg ggccgagatc cgttgatctt cctgcatccg ccagaggcgg
     4921 gatgcgaaga atgcgatgcc gctcgccagt cgattggctg agctcatgag cggagaacga
     4981 gatgacgttg gaggggcaag gtcgcgctga ttgctggggc aacacgtgga gcggatcggg
     5041 gattgtcttt cttcagctcg ctgatgatat gctgacgctc aatgccgttt ggcctccgac
     5101 taacgaaaat cccgcatttg gacggctgat ccatggtact cccgttgaat atggctcata
     5161 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     5221 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttc cccccccccg
     5281 acgtcaggtg gctagctggc gggaacggca atgctcctgg cggcggcggg gctggtgccc
     5341 aggtctccgc acagagcggc ggtgctggcg ctcgcggcca ggcgtggttc ttcgcgtact
     5401 gacaagaaac ccccctcttt aggactcagt gtccttggga ggggggcttt ttgcgtttca
     5461 ggaggtcttg gccaggcatg ccacggatgc caccacaggc actacaacgg agttcgccac
     5521 gtacatcacc acaaccaccg attctggcgg tgagctcccc gatattcagc ggaaatggct
     5581 tggtatcgac caagattcgt agaaccccgt ctcgtctggc tggtattcaa aacgggcgca
     5641 acgaaacacg caacgagaca ggcatggccc aaaccagaaa actagcgtct accaggactt
     5701 ttacctgtcc gacccgttgc aacggaaccc cccacggaac ccccgcgaca cccgctcccc
     5761 aattgcgtta gaacagcggt ggattgtcgg cttcgttgtg ggccttttga gccgcttcct
     5821 gttctgccgc acgctctttc ctcgcccgat agccgagtcg cttaacggtg tccagatgca
     5881 gcccgaaatg tttggccgtt tgcggccaag agtggccctc gtcgtcgtga taggcgcgga
     5941 tgcgttcgcg gcgtgcagcc tgctcggcga gccactcgct gcgttcctgc gccacgagcc
     6001 ggacgacgtg gcgttcggat agtccggtga ttcgagcgcc ttcggcggcg gtcacgcgcc
     6061 gctttttgcg gacagtcggc tgccggttgt agccgtcgct gtagccgtcg ctcatagcaa
     6121 tgcctccatg catggctgac gcggactttg cgcgccgcgc aactgtgctc gccgccgtgc
     6181 gcgctgctgc gcccttccgc gagatggccg actggcgcgc actgagtgtg gcctcgtaga
     6241 ccacgatccc gtccgcccaa atgcgcgact tggttgtgat ccaacgccaa atgctgttgg
     6301 cgatggcgcg gacctcgctg tccggtagcg gtccgggaca cacgtcgttg cacggaaatt
     6361 cggcgtttcg cgcgtggcac tcggcataga tcgcgcggcc gagtccgtcc acgttccggg
     6421 tcggcaggta gatccgcatg agggcgggac gataggccca caacctgacg gaatcgaaca
     6481 gtgcgcaatt ccgccctagc ggcgtcggag ccgctttgta cgtggtctgc tgacgccagc
     6541 gcggcggtgg catgttcgcg ccgagctcgg cctcgatgtg gctgagtgtg tagagatctg
     6601 agtggagcca ttccgtttcc caggcgatgt ggccggggtt tttggtcatg aggcctgagt
     6661 aactgcggtc gccatcgacg gcgcgccgaa ggccttcggc gcacgccgcc atgtatgcga
     6721 gcggcttacg ccgcgcgtat tcggtgcgtg gaacaggggc gttgagtgcc cacactgcgt
     6781 gtgcgtggcc gttggcgcga ttgcccacga tcgcgttggg cagcggatgg gacccccggg
     6841 cgctgagcgc tcggagcgct gcgtctggat ggtctacgtc cacgaccagc aggtttgcca
     6901 gcgctgttgg gttcgcctcg atgtaccggc ggcctagggc cgacgcgcgg ctttggcggt
     6961 agatcccctc gagcagatcg tcgcttgcca gcggccagta cggcagccag agctgctcaa
     7021 attcgtcggc gacgtggctc acgcttggta gtagaccacg attaatcacc ggtgtatggt
     7081 ccgacacgag ctccaagtca gatatttcgc tgaggggcca ccccacaact gcacactccc
     7141 ccgctctccc gtcgagccct ggtggtggaa caccagcgac agccgagcac ccccaaccac
     7201 ctgtaccaac caggaggaac acatgcgtcg tttcgaggac gtttccgggc cgctgagagc
     7261 cgctgtggcg gccgtacacg ccgccttaga cccgttagac cccctgccgc ctgaatgcgc
     7321 gggtacgagc cacacagcgc ccgaacttac ggagctggtg ggctcactct agatcacggc
     7381 tctagctgat caccgcggcc atgatggcat aaaacgaaag gcccagtctt tcgactgagc
     7441 ctttcgtttt atttgatgcc tggcagtcga tcgtacgcta gttaactacg tcgacatcga
     7501 tttaattaat atgcatcaat tgatttaaat actagtgtta actatttaat tggggaccct
     7561 agaggtcccc ttttttattt taaaaatttt ttcaagggga cctctagggt ccccagctgg
     7621 ctagcgagtc atgaggtcac catgttcttt cctgcgttat cccctgattc tgtggataac
     7681 cgtattaccg cctttgagtg agctgatacc agcggataac aatttcacac aggaaacagc
     7741 tatgaccatg attacgccaa gctatcaact ttgtatagaa aagttgcatg accaacttcg
     7801 ataacgttct cggctcgatg atccccgggg gccccccctc gaggtcgacg gtatcgattt
     7861 aattaatatg catcaattga tttaaatact agtgttaact atttaattgg ggaccctaga
     7921 ggtccccttt tttattttaa aaattttttc aaggggacct ctagggtccc cagctggcta
     7981 gcgagtcatg aggtcaccat ggtcattagg agccgctctc gcacttcagc tgcttctcca
     8041 agccgcagat gatcagctcc aggccgaaca agaaggccgg ctcggcgccc tggtggtcga
     8101 acagctcgat ggcctggcgc agcaacggcg gcatgctgtc ggtggtcggg gtctcgcgct
     8161 cctccttggc gacctggtgc tcctggtcct ccaacacgca gcccagggtg aagtggccca
     8221 cggccgacag ggcgtacaac gcgttctcca gcgagaagcc ctgctggcac aggaaggcca
     8281 actggttctc cagggtctcg tactgcttct cggtcggccg ggtgcccagg tgcaccttgg
     8341 cgccgtcgcg gtggctcagc aaggcgcagc ggaagctctt ggcgttgttg cgcaggaagt
     8401 cctgccagct ctcgccctcc agcgggcaga agtgggtgtg gtggcggtcc aacatctcga
     8461 tggccagggc gtcgagcagg gcccgcttgt tcttgacgtg ccagtacagg gtcggctgct
     8521 cgacgcccag cttctgggcc agcttgcggg tggtcaggcc ctcgatgccg acctcgttca
     8581 gcagctccag ggcggagttg atgaccttgg acttgtccag gcgggacatt gcgaagtgat
     8641 tcctcctgat ggtaccggat ccgtggcagg agggtcgcct gcgtcggcaa ctcgcccagg
     8701 ctacacgaga gccctcgact gcgaaaatgc cgaccaggca ccaccgcact ccaggcttaa
     8761 cccggtgcga cgatccaata ttacgagatc tgacaagttt gtacaaaaaa gcaggctacc
     8821 aggcctagat ctggggaccc tagaggtccc cttttttatt ttaaaaattt tttcacgtgc
     8881 cagccgattg gactagaaat attgga
//
