LOCUS       sequence-211574-        8559 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 8559)
  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 8559)
  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 (Kan)
FEATURES             Location/Qualifiers
     source          1..8559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(307..331)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(410..715)
                     /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(512..531)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1057..1716)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1631..1650
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1770..1776,1777..1794,1795..1800))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1825..1949)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1968..1988)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1992..2012)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2039..2062
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2069..2779
                     /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(2096..2124)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2719..2739
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2786..2815
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     rep_origin      complement(3085..3673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3165..3184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4004..4819)
                     /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     4727..4746
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4811..4830
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(7069..7115)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7371..7393
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7390..7406
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7390..7406
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    7426..7446
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7499..7515
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7500..7516
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8453..8477
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tattggatcg tcggcaccgt cacggccgtg ggaggcggca cgatccgcga cgtgatgatc
       61 ggccgcatcc ccacggtgct gcgcagtgag ctctacgcca tcccggcgtt gatctgtgcg
      121 ttcgcacgca caggcccggt gtgagaaggg tctctgacga gcgggagaac tccctatcag
      181 tgatagagtt tgtcctccct atcagtgata gataggctct gggagtaccc gtgtgtacga
      241 ccagcacggc atacatcatt tcgacgccga gagattcgcc gcccgaaatg agcacgatcc
      301 gcatgcaccc agctttcttg tacaaagtgg gctcgaggca tgcgaaggag atatacatat
      361 gagttgaaga gcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc
      421 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
      481 cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
      541 catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg
      601 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
      661 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
      721 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
      781 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
      841 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
      901 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
      961 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
     1021 ctgttatctg gcttttagta agccggatcc acgcgtttac gccccgccct gccactcatc
     1081 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     1141 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     1201 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     1261 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     1321 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     1381 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     1441 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     1501 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     1561 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     1621 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     1681 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     1741 tctcgacgga tcctaactca aaatccacac attatacgag ccggaagcat aaagtgtaaa
     1801 gcctggggtg cctaatgcgg ccgccatagt gactggatat gttgtgtttt acagtattat
     1861 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     1921 tctcgttcag cttttttgta caaacttgtg atatcgaatt cgctagctac ctttctcttc
     1981 ttttttggat ctacctttct cttctttttt ggggatccac tagttcagct cttctgcatt
     2041 agaagttttg tttcaaggtc cacaattcag caaaggagaa gaacttttca ctggagttgt
     2101 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     2161 gggtgaaggt gatgctacat acggaaaact cacccttaaa tttatttgca ctactggaaa
     2221 actacctgtt ccttggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc
     2281 ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     2341 tgtacaggaa cgcactatat ctttcaaaga tgacgggaac tacaagacgc gtgctgaagt
     2401 caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga
     2461 agatggaaac attctcggac acaaactaga gtacaactat aactcacaca atgtatacat
     2521 cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacattga
     2581 agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     2641 tgtcctttta ccagacaacc attacctgtc gacacaatct gccctttcga aagatcccaa
     2701 cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg
     2761 catggatgag ctctacaaaa ctagtcatca tcatcaccat catcatcacc atcattaatt
     2821 atacatagtt gcttaagctg caggggaccc tagaggtccc cttttttatt ttaaaaattt
     2881 tttataaccg gtaagaagtc caacacgccg ctcctccccg atgttgcgta gccgcttcaa
     2941 gcttaagcaa cggccccagt gagtagcgac tttggatccg gtacccgcaa cggtcaacca
     3001 gcgtctacgc gtttcactgg ccgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3061 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3121 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3181 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3241 gtccgccttt ctcccctcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3301 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3361 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3421 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3481 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3541 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3601 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3661 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc atggaactag
     3721 tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat cgccccatca
     3781 tccagccaga aagtgaggga gccacggttg gtgagagctt tgttgtaggt ggaccagttg
     3841 gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc
     3901 tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc cgtcaagtca
     3961 gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     4021 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     4081 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     4141 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     4201 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     4261 gcaaaagatt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     4321 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     4381 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     4441 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     4501 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     4561 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     4621 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatggg
     4681 gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     4741 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     4801 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     4861 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     4921 cgtggctttc cccccccccg acgtcaggtg gctagctggc gggaacggca atgctcctgg
     4981 cggcggcggg gctggtgccc aggtctccgc acagagcggc ggtgctggcg ctcgcggcca
     5041 ggcgtggttc ttcgcgtact gacaagaaac ccccctcttt aggactcagt gtccttggga
     5101 ggggggcttt ttgcgtttca ggaggtcttg gccaggcatg ccacggatgc caccacaggc
     5161 actacaacgg agttcgccac gtacatcacc acaaccaccg attctggcgg tgagctcccc
     5221 gatattcagc ggaaatggct tggtatcgac caagattcgt agaaccccgt ctcgtctggc
     5281 tggtattcaa aacgggcgca acgaaacacg caacgagaca ggcatggccc aaaccagaaa
     5341 actagcgtct accaggactt ttacctgtcc gacccgttgc aacggaaccc cccacggaac
     5401 ccccgcgaca cccgctcccc aattgcgtta gaacagcggt ggattgtcgg cttcgttgtg
     5461 ggccttttga gccgcttcct gttctgccgc acgctctttc ctcgcccgat agccgagtcg
     5521 cttaacggtg tccagatgca gcccgaaatg tttggccgtt tgcggccaag agtggccctc
     5581 gtcgtcgtga taggcgcgga tgcgttcgcg gcgtgcagcc tgctcggcga gccactcgct
     5641 gcgttcctgc gccacgagcc ggacgacgtg gcgttcggat agtccggtga ttcgagcgcc
     5701 ttcggcggcg gtcacgcgcc gctttttgcg gacagtcggc tgccggttgt agccgtcgct
     5761 gtagccgtcg ctcatagcaa tgcctccatg catggctgac gcggactttg cgcgccgcgc
     5821 aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc gagatggccg actggcgcgc
     5881 actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa atgcgcgact tggttgtgat
     5941 ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg tccggtagcg gtccgggaca
     6001 cacgtcgttg cacggaaatt cggcgtttcg cgcgtggcac tcggcataga tcgcgcggcc
     6061 gagtccgtcc acgttccggg tcggcaggta gatccgcatg agggcgggac gataggccca
     6121 caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc ggcgtcggag ccgctttgta
     6181 cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg ccgagctcgg cctcgatgtg
     6241 gctgagtgtg tagagatctg agtggagcca ttccgtttcc caggcgatgt ggccggggtt
     6301 tttggtcatg aggcctgagt aactgcggtc gccatcgacg gcgcgccgaa ggccttcggc
     6361 gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat tcggtgcgtg gaacaggggc
     6421 gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga ttgcccacga tcgcgttggg
     6481 cagcggatgg gacccccggg cgctgagcgc tcggagcgct gcgtctggat ggtctacgtc
     6541 cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg atgtaccggc ggcctagggc
     6601 cgacgcgcgg ctttggcggt agatcccctc gagcagatcg tcgcttgcca gcggccagta
     6661 cggcagccag agctgctcaa attcgtcggc gacgtggctc acgcttggta gtagaccacg
     6721 attaatcacc ggtgtatggt ccgacacgag ctccaagtca gatatttcgc tgaggggcca
     6781 ccccacaact gcacactccc ccgctctccc gtcgagccct ggtggtggaa caccagcgac
     6841 agccgagcac ccccaaccac ctgtaccaac caggaggaac acatgcgtcg tttcgaggac
     6901 gtttccgggc cgctgagagc cgctgtggcg gccgtacacg ccgccttaga cccgttagac
     6961 cccctgccgc ctgaatgcgc gggtacgagc cacacagcgc ccgaacttac ggagctggtg
     7021 ggctcactct agatcacggc tctagctgat caccgcggcc atgatggcat aaaacgaaag
     7081 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggcagtcga tcgtacgcta
     7141 gttaactacg tcgacatcga tttaattaat atgcatcaat tgatttaaat actagtgtta
     7201 actatttaat tggggaccct agaggtcccc ttttttattt taaaaatttt ttcaagggga
     7261 cctctagggt ccccagctgg ctagcgagtc atgaggtcac catgttcttt cctgcgttat
     7321 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc agcggataac
     7381 aatttcacac aggaaacagc tatgaccatg attacgccaa gctatcaact ttgtatagaa
     7441 aagttgcatg accaacttcg ataacgttct cggctcgatg atccccgggg gccccccctc
     7501 gaggtcgacg gtatcgattt aattaatatg catcaattga tttaaatact agtgttaact
     7561 atttaattgg ggaccctaga ggtccccttt tttattttaa aaattttttc aaggggacct
     7621 ctagggtccc cagctggcta gcgagtcatg aggtcaccat ggtcattagg agccgctctc
     7681 gcacttcagc tgcttctcca agccgcagat gatcagctcc aggccgaaca agaaggccgg
     7741 ctcggcgccc tggtggtcga acagctcgat ggcctggcgc agcaacggcg gcatgctgtc
     7801 ggtggtcggg gtctcgcgct cctccttggc gacctggtgc tcctggtcct ccaacacgca
     7861 gcccagggtg aagtggccca cggccgacag ggcgtacaac gcgttctcca gcgagaagcc
     7921 ctgctggcac aggaaggcca actggttctc cagggtctcg tactgcttct cggtcggccg
     7981 ggtgcccagg tgcaccttgg cgccgtcgcg gtggctcagc aaggcgcagc ggaagctctt
     8041 ggcgttgttg cgcaggaagt cctgccagct ctcgccctcc agcgggcaga agtgggtgtg
     8101 gtggcggtcc aacatctcga tggccagggc gtcgagcagg gcccgcttgt tcttgacgtg
     8161 ccagtacagg gtcggctgct cgacgcccag cttctgggcc agcttgcggg tggtcaggcc
     8221 ctcgatgccg acctcgttca gcagctccag ggcggagttg atgaccttgg acttgtccag
     8281 gcgggacatt gcgaagtgat tcctcctgat ggtaccggat ccgtggcagg agggtcgcct
     8341 gcgtcggcaa ctcgcccagg ctacacgaga gccctcgact gcgaaaatgc cgaccaggca
     8401 ccaccgcact ccaggcttaa cccggtgcga cgatccaata ttacgagatc tgacaagttt
     8461 gtacaaaaaa gcaggctacc aggcctagat ctggggaccc tagaggtccc cttttttatt
     8521 ttaaaaattt tttcacgtgc cagccgattg gactagaaa
//
