LOCUS       sequence-211279-        9460 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 9460)
  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 9460)
  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 (Hyg)
FEATURES             Location/Qualifiers
     source          1..9460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(277..301)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(380..685)
                     /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(482..501)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1027..1686)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1601..1620
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1740..1746,1747..1764,1765..1770))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1795..1919)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1938..1958)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1962..1982)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2009..2032
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2039..2749
                     /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(2066..2094)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2689..2709
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2756..2785
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     rep_origin      complement(3055..3643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3135..3154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5682..5701
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(7940..7986)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     8242..8264
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8261..8277
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8261..8277
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    8297..8317
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     8370..8386
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8371..8387
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9324..9348
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ggaggcggca cgatccgcga cgtgatgatc ggccgcatcc ccacggtgct gcgcagtgag
       61 ctctacgcca tcccggcgtt gatctgtgcg ttcgcacgca caggcccggt gtgagaaggg
      121 tctctgacga gcgggagaac tccctatcag tgatagagtt tgtcctccct atcagtgata
      181 gataggctct gggagtaccc gtgtgtacga ccagcacggc atacatcatt tcgacgccga
      241 gagattcgcc gcccgaaatg agcacgatcc gcatgcaccc agctttcttg tacaaagtgg
      301 gctcgaggca tgcgaaggag atatacatat gagttgaaga gcgacctgca gactggctgt
      361 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
      421 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
      481 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
      541 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
      601 cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
      661 tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg
      721 ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt
      781 tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg
      841 acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct
      901 gcttcatagc atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat
      961 accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc
     1021 acgcgtttac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1081 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1141 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     1201 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1261 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1321 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1381 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1441 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     1501 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1561 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1621 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1681 ctccatttta gcttccttag ctcctgaaaa tctcgacgga tcctaactca aaatccacac
     1741 attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt
     1801 gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
     1861 aatatattga tatttatatc attttacgtt tctcgttcag cttttttgta caaacttgtg
     1921 atatcgaatt cgctagctac ctttctcttc ttttttggat ctacctttct cttctttttt
     1981 ggggatccac tagttcagct cttctgcatt agaagttttg tttcaaggtc cacaattcag
     2041 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     2101 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgctacat acggaaaact
     2161 cacccttaaa tttatttgca ctactggaaa actacctgtt ccttggccaa cacttgtcac
     2221 tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga
     2281 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
     2341 tgacgggaac tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
     2401 tatcgagtta aagggtattg attttaaaga agatggaaac attctcggac acaaactaga
     2461 gtacaactat aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
     2521 agctaacttc aaaattcgcc acaacattga agatggttcc gttcaactag cagaccatta
     2581 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     2641 gacacaatct gccctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
     2701 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaaa ctagtcatca
     2761 tcatcaccat catcatcacc atcattaatt atacatagtt gcttaagctg caggggaccc
     2821 tagaggtccc cttttttatt ttaaaaattt tttataaccg gtaagaagtc caacacgccg
     2881 ctcctccccg atgttgcgta gccgcttcaa gcttaagcaa cggccccagt gagtagcgac
     2941 tttggatccg gtacccgcaa cggtcaacca gcgtctacgc gtttcactgg ccgacatgtg
     3001 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3061 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3121 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3181 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccctcgg gaagcgtggc
     3241 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3301 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3361 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3421 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3481 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3541 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3601 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3661 ttctacgggg tctgacgctc atggaactag tgaggtctgc ctcgtgaaga aggtgttgct
     3721 gactcatacc aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg
     3781 gtgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa
     3841 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt
     3901 attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat
     3961 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaag tattctagag
     4021 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4081 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4141 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4201 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gacgtggccg accagcccgt
     4261 catcgtcaac gcctgatccg cggtgcggac aggccgtgtc gcgaccggcc gtgcggaatt
     4321 aagccggccc gtaccctgtg aatagaggtc cgctgtgaca caagaatccc tgttacttct
     4381 cgaccgtatt gattcggatg attcctacgc gagcctgcgg aacgaccagg aattctggga
     4441 gccgctggcc cgccgagccc tggaggagct cgggctgccg gtgccgccgg tgctgcgggt
     4501 gcccggcgag agcaccaacc ccgtactggt cggcgagccc ggcccggtga tcaagctgtt
     4561 cggcgagcac tggtgcggtc cggagagcct cgcgtcggag tcggaggcgt acgcggtcct
     4621 ggcggacgcc ccggtgccgg tgccccgcct cctcggccgc ggcgagctgc ggcccggcac
     4681 cggagcctgg ccgtggccct acctggtgat gagccggatg accggcacca cctggcggtc
     4741 cgcgatggac ggcacgaccg accggaacgc gctgctcgcc ctggcccgcg aactcggccg
     4801 ggtgctcggc cggctgcaca gggtgccgct gaccgggaac accgtgctca ccccccattc
     4861 cgaggtcttc ccggaactgc tgcgggaacg ccgcgcggcg accgtcgagg accaccgcgg
     4921 gtggggctac ctctcgcccc ggctgctgga ccgcctggag gactggctgc cggacgtgga
     4981 cacgctgctg gccggccgcg aaccccggtt cgtccacggc gacctgcacg ggaccaacat
     5041 cttcgtggac ctggccgcga ccgaggtcac cgggatcgtc gacttcaccg acgtctatgc
     5101 gggagactcc cgctacagcc tggtgcaact gcatctcaac gccttccggg gcgaccgcga
     5161 gatcctggcc gcgctgctcg acggggcgca gtggaagcgg accgaggact tcgcccgcga
     5221 actgctcgcc ttcaccttcc tgcacgactt cgaggtgttc gaggagaccc cgctggatct
     5281 ctccggcttc accgatccgg aggaactggc gcagttcctc tgggggccgc cggacaccgc
     5341 ccccggcccc cggcggccgc cgggcggagc cccgcccggc ggcccgggag ccccgggccc
     5401 gcgccgaagc ccgctgctgc gagcccggag cgggccggcc gacggcggtg cgggcccggc
     5461 ggcggacgct cagcagcggc gggcgtgaaa ggccctggca tcctcgatca tctcctccag
     5521 ggtgggtcgg ccgggcttcc atcccagctc ggcaaggatc aacgaggaca gtcgcacgac
     5581 gaagttcttc tggatcgcgc ccgtgctgga agcactcaac ctcgaagcgt gtggttgcgg
     5641 agccatctag caaccacacg accatggtac tcccgttgaa tatggctcat aacacccctt
     5701 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     5761 gcaatgtaac atcagagatt ttgagacaca acgtggcttt cccccccccc gacgtcaggt
     5821 ggctagctgg cgggaacggc aatgctcctg gcggcggcgg ggctggtgcc caggtctccg
     5881 cacagagcgg cggtgctggc gctcgcggcc aggcgtggtt cttcgcgtac tgacaagaaa
     5941 cccccctctt taggactcag tgtccttggg aggggggctt tttgcgtttc aggaggtctt
     6001 ggccaggcat gccacggatg ccaccacagg cactacaacg gagttcgcca cgtacatcac
     6061 cacaaccacc gattctggcg gtgagctccc cgatattcag cggaaatggc ttggtatcga
     6121 ccaagattcg tagaaccccg tctcgtctgg ctggtattca aaacgggcgc aacgaaacac
     6181 gcaacgagac aggcatggcc caaaccagaa aactagcgtc taccaggact tttacctgtc
     6241 cgacccgttg caacggaacc ccccacggaa cccccgcgac acccgctccc caattgcgtt
     6301 agaacagcgg tggattgtcg gcttcgttgt gggccttttg agccgcttcc tgttctgccg
     6361 cacgctcttt cctcgcccga tagccgagtc gcttaacggt gtccagatgc agcccgaaat
     6421 gtttggccgt ttgcggccaa gagtggccct cgtcgtcgtg ataggcgcgg atgcgttcgc
     6481 ggcgtgcagc ctgctcggcg agccactcgc tgcgttcctg cgccacgagc cggacgacgt
     6541 ggcgttcgga tagtccggtg attcgagcgc cttcggcggc ggtcacgcgc cgctttttgc
     6601 ggacagtcgg ctgccggttg tagccgtcgc tgtagccgtc gctcatagca atgcctccat
     6661 gcatggctga cgcggacttt gcgcgccgcg caactgtgct cgccgccgtg cgcgctgctg
     6721 cgcccttccg cgagatggcc gactggcgcg cactgagtgt ggcctcgtag accacgatcc
     6781 cgtccgccca aatgcgcgac ttggttgtga tccaacgcca aatgctgttg gcgatggcgc
     6841 ggacctcgct gtccggtagc ggtccgggac acacgtcgtt gcacggaaat tcggcgtttc
     6901 gcgcgtggca ctcggcatag atcgcgcggc cgagtccgtc cacgttccgg gtcggcaggt
     6961 agatccgcat gagggcggga cgataggccc acaacctgac ggaatcgaac agtgcgcaat
     7021 tccgccctag cggcgtcgga gccgctttgt acgtggtctg ctgacgccag cgcggcggtg
     7081 gcatgttcgc gccgagctcg gcctcgatgt ggctgagtgt gtagagatct gagtggagcc
     7141 attccgtttc ccaggcgatg tggccggggt ttttggtcat gaggcctgag taactgcggt
     7201 cgccatcgac ggcgcgccga aggccttcgg cgcacgccgc catgtatgcg agcggcttac
     7261 gccgcgcgta ttcggtgcgt ggaacagggg cgttgagtgc ccacactgcg tgtgcgtggc
     7321 cgttggcgcg attgcccacg atcgcgttgg gcagcggatg ggacccccgg gcgctgagcg
     7381 ctcggagcgc tgcgtctgga tggtctacgt ccacgaccag caggtttgcc agcgctgttg
     7441 ggttcgcctc gatgtaccgg cggcctaggg ccgacgcgcg gctttggcgg tagatcccct
     7501 cgagcagatc gtcgcttgcc agcggccagt acggcagcca gagctgctca aattcgtcgg
     7561 cgacgtggct cacgcttggt agtagaccac gattaatcac cggtgtatgg tccgacacga
     7621 gctccaagtc agatatttcg ctgaggggcc accccacaac tgcacactcc cccgctctcc
     7681 cgtcgagccc tggtggtgga acaccagcga cagccgagca cccccaacca cctgtaccaa
     7741 ccaggaggaa cacatgcgtc gtttcgagga cgtttccggg ccgctgagag ccgctgtggc
     7801 ggccgtacac gccgccttag acccgttaga ccccctgccg cctgaatgcg cgggtacgag
     7861 ccacacagcg cccgaactta cggagctggt gggctcactc tagatcacgg ctctagctga
     7921 tcaccgcggc catgatggca taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     7981 tatttgatgc ctggcagtcg atcgtacgct agttaactac gtcgacatcg atttaattaa
     8041 tatgcatcaa ttgatttaaa tactagtgtt aactatttaa ttggggaccc tagaggtccc
     8101 cttttttatt ttaaaaattt tttcaagggg acctctaggg tccccagctg gctagcgagt
     8161 catgaggtca ccatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8221 gcctttgagt gagctgatac cagcggataa caatttcaca caggaaacag ctatgaccat
     8281 gattacgcca agctatcaac tttgtataga aaagttgcat gaccaacttc gataacgttc
     8341 tcggctcgat gatccccggg ggccccccct cgaggtcgac ggtatcgatt taattaatat
     8401 gcatcaattg atttaaatac tagtgttaac tatttaattg gggaccctag aggtcccctt
     8461 ttttatttta aaaatttttt caaggggacc tctagggtcc ccagctggct agcgagtcat
     8521 gaggtcacca tggtcattag gagccgctct cgcacttcag ctgcttctcc aagccgcaga
     8581 tgatcagctc caggccgaac aagaaggccg gctcggcgcc ctggtggtcg aacagctcga
     8641 tggcctggcg cagcaacggc ggcatgctgt cggtggtcgg ggtctcgcgc tcctccttgg
     8701 cgacctggtg ctcctggtcc tccaacacgc agcccagggt gaagtggccc acggccgaca
     8761 gggcgtacaa cgcgttctcc agcgagaagc cctgctggca caggaaggcc aactggttct
     8821 ccagggtctc gtactgcttc tcggtcggcc gggtgcccag gtgcaccttg gcgccgtcgc
     8881 ggtggctcag caaggcgcag cggaagctct tggcgttgtt gcgcaggaag tcctgccagc
     8941 tctcgccctc cagcgggcag aagtgggtgt ggtggcggtc caacatctcg atggccaggg
     9001 cgtcgagcag ggcccgcttg ttcttgacgt gccagtacag ggtcggctgc tcgacgccca
     9061 gcttctgggc cagcttgcgg gtggtcaggc cctcgatgcc gacctcgttc agcagctcca
     9121 gggcggagtt gatgaccttg gacttgtcca ggcgggacat tgcgaagtga ttcctcctga
     9181 tggtaccgga tccgtggcag gagggtcgcc tgcgtcggca actcgcccag gctacacgag
     9241 agccctcgac tgcgaaaatg ccgaccaggc accaccgcac tccaggctta acccggtgcg
     9301 acgatccaat attacgagat ctgacaagtt tgtacaaaaa agcaggctac caggcctaga
     9361 tctggggacc ctagaggtcc ccttttttat tttaaaaatt ttttcacgtg ccagccgatt
     9421 ggactagaaa tattggatcg tcggcaccgt cacggccgtg
//
