LOCUS       sequence-211290-        7836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pMINTC3H, but containing oriM for episomal replication 
            in mycobacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  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 7836)
  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: pMEXC3H
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(353..377)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(456..761)
                     /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(558..577)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1103..1762)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1677..1696
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1816..1822,1823..1840,1841..1846))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1871..1995)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2014..2034)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2038..2058)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2085..2108
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2109..2138
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     rep_origin      complement(2408..2996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2488..2507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3327..4142)
                     /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     4050..4069
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4134..4153
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(6392..6438)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6694..6716
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6713..6729
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6713..6729
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6749..6769
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     6822..6838
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6823..6839
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7776..7800
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttttatttta aaaatttttt cacgtgccag ccgattggac tagaaatatt ggatcgtcgg
       61 caccgtcacg gccgtgggag gcggcacgat ccgcgacgtg atgatcggcc gcatccccac
      121 ggtgctgcgc agtgagctct acgccatccc ggcgttgatc tgtgcgttcg cacgcacagg
      181 cccggtgtga gaagggtctc tgacgagcgg gagaactccc tatcagtgat agagtttgtc
      241 ctccctatca gtgatagata ggctctggga gtacccgtgt gtacgaccag cacggcatac
      301 atcatttcga cgccgagaga ttcgccgccc gaaatgagca cgatccgcat gcacccagct
      361 ttcttgtaca aagtgggctc gaggcatgcg aaggagatat acatatgagt tgaagagcga
      421 cctgcagact ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta
      481 atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg
      541 gagaccggca cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc
      601 accaccgggt aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg
      661 atcaccatcc gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga
      721 taacggctct ctcttttata ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc
      781 gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt
      841 ttccgctttc cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac
      901 cagaccggag atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact
      961 gtcactgtaa tacgctgctt catagcatac ctctttttga catacttcgg gtatacatat
     1021 cagtatatat tcttataccg caaaaatcag cgcgcaaata cgcatactgt tatctggctt
     1081 ttagtaagcc ggatccacgc gtttacgccc cgccctgcca ctcatcgcag tactgttgta
     1141 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     1201 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     1261 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     1321 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     1381 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     1441 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     1501 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat
     1561 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     1621 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     1681 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     1741 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gacggatcct
     1801 aactcaaaat ccacacatta tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1861 atgcggccgc catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt
     1921 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt
     1981 tttgtacaaa cttgtgatat cgaattcgct agctaccttt ctcttctttt ttggatctac
     2041 ctttctcttc ttttttgggg atccactagt tcagctcttc tgcattagaa gttttgtttc
     2101 aaggtccaca tcatcatcac catcatcatc accatcatta attatacata gttgcttaag
     2161 ctgcagggga ccctagaggt cccctttttt attttaaaaa ttttttataa ccggtaagaa
     2221 gtccaacacg ccgctcctcc ccgatgttgc gtagccgctt caagcttaag caacggcccc
     2281 agtgagtagc gactttggat ccggtacccg caacggtcaa ccagcgtcta cgcgtttcac
     2341 tggccgacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctcccct
     2581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2821 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     2881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3001 atcctttgat cttttctacg gggtctgacg ctcatggaac tagtgaggtc tgcctcgtga
     3061 agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag
     3121 ggagccacgg ttggtgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg
     3181 ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc
     3241 aaaagttcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat gctctgccag
     3301 tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc
     3361 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     3421 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     3481 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     3541 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag attatgcatt
     3601 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     3661 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     3721 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     3781 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     3841 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     3901 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     3961 acgctacctt tgccatgttt cagaaacaac tctggcgcat ggggcttccc atacaagcga
     4021 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     4081 gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg aatatggctc
     4141 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     4201 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct ttcccccccc
     4261 ccgacgtcag gtggctagct ggcgggaacg gcaatgctcc tggcggcggc ggggctggtg
     4321 cccaggtctc cgcacagagc ggcggtgctg gcgctcgcgg ccaggcgtgg ttcttcgcgt
     4381 actgacaaga aacccccctc tttaggactc agtgtccttg ggaggggggc tttttgcgtt
     4441 tcaggaggtc ttggccaggc atgccacgga tgccaccaca ggcactacaa cggagttcgc
     4501 cacgtacatc accacaacca ccgattctgg cggtgagctc cccgatattc agcggaaatg
     4561 gcttggtatc gaccaagatt cgtagaaccc cgtctcgtct ggctggtatt caaaacgggc
     4621 gcaacgaaac acgcaacgag acaggcatgg cccaaaccag aaaactagcg tctaccagga
     4681 cttttacctg tccgacccgt tgcaacggaa ccccccacgg aacccccgcg acacccgctc
     4741 cccaattgcg ttagaacagc ggtggattgt cggcttcgtt gtgggccttt tgagccgctt
     4801 cctgttctgc cgcacgctct ttcctcgccc gatagccgag tcgcttaacg gtgtccagat
     4861 gcagcccgaa atgtttggcc gtttgcggcc aagagtggcc ctcgtcgtcg tgataggcgc
     4921 ggatgcgttc gcggcgtgca gcctgctcgg cgagccactc gctgcgttcc tgcgccacga
     4981 gccggacgac gtggcgttcg gatagtccgg tgattcgagc gccttcggcg gcggtcacgc
     5041 gccgcttttt gcggacagtc ggctgccggt tgtagccgtc gctgtagccg tcgctcatag
     5101 caatgcctcc atgcatggct gacgcggact ttgcgcgccg cgcaactgtg ctcgccgccg
     5161 tgcgcgctgc tgcgcccttc cgcgagatgg ccgactggcg cgcactgagt gtggcctcgt
     5221 agaccacgat cccgtccgcc caaatgcgcg acttggttgt gatccaacgc caaatgctgt
     5281 tggcgatggc gcggacctcg ctgtccggta gcggtccggg acacacgtcg ttgcacggaa
     5341 attcggcgtt tcgcgcgtgg cactcggcat agatcgcgcg gccgagtccg tccacgttcc
     5401 gggtcggcag gtagatccgc atgagggcgg gacgataggc ccacaacctg acggaatcga
     5461 acagtgcgca attccgccct agcggcgtcg gagccgcttt gtacgtggtc tgctgacgcc
     5521 agcgcggcgg tggcatgttc gcgccgagct cggcctcgat gtggctgagt gtgtagagat
     5581 ctgagtggag ccattccgtt tcccaggcga tgtggccggg gtttttggtc atgaggcctg
     5641 agtaactgcg gtcgccatcg acggcgcgcc gaaggccttc ggcgcacgcc gccatgtatg
     5701 cgagcggctt acgccgcgcg tattcggtgc gtggaacagg ggcgttgagt gcccacactg
     5761 cgtgtgcgtg gccgttggcg cgattgccca cgatcgcgtt gggcagcgga tgggaccccc
     5821 gggcgctgag cgctcggagc gctgcgtctg gatggtctac gtccacgacc agcaggtttg
     5881 ccagcgctgt tgggttcgcc tcgatgtacc ggcggcctag ggccgacgcg cggctttggc
     5941 ggtagatccc ctcgagcaga tcgtcgcttg ccagcggcca gtacggcagc cagagctgct
     6001 caaattcgtc ggcgacgtgg ctcacgcttg gtagtagacc acgattaatc accggtgtat
     6061 ggtccgacac gagctccaag tcagatattt cgctgagggg ccaccccaca actgcacact
     6121 cccccgctct cccgtcgagc cctggtggtg gaacaccagc gacagccgag cacccccaac
     6181 cacctgtacc aaccaggagg aacacatgcg tcgtttcgag gacgtttccg ggccgctgag
     6241 agccgctgtg gcggccgtac acgccgcctt agacccgtta gaccccctgc cgcctgaatg
     6301 cgcgggtacg agccacacag cgcccgaact tacggagctg gtgggctcac tctagatcac
     6361 ggctctagct gatcaccgcg gccatgatgg cataaaacga aaggcccagt ctttcgactg
     6421 agcctttcgt tttatttgat gcctggcagt cgatcgtacg ctagttaact acgtcgacat
     6481 cgatttaatt aatatgcatc aattgattta aatactagtg ttaactattt aattggggac
     6541 cctagaggtc ccctttttta ttttaaaaat tttttcaagg ggacctctag ggtccccagc
     6601 tggctagcga gtcatgaggt caccatgttc tttcctgcgt tatcccctga ttctgtggat
     6661 aaccgtatta ccgcctttga gtgagctgat accagcggat aacaatttca cacaggaaac
     6721 agctatgacc atgattacgc caagctatca actttgtata gaaaagttgc atgaccaact
     6781 tcgataacgt tctcggctcg atgatccccg ggggcccccc ctcgaggtcg acggtatcga
     6841 tttaattaat atgcatcaat tgatttaaat actagtgtta actatttaat tggggaccct
     6901 agaggtcccc ttttttattt taaaaatttt ttcaagggga cctctagggt ccccagctgg
     6961 ctagcgagtc atgaggtcac catggtcatt aggagccgct ctcgcacttc agctgcttct
     7021 ccaagccgca gatgatcagc tccaggccga acaagaaggc cggctcggcg ccctggtggt
     7081 cgaacagctc gatggcctgg cgcagcaacg gcggcatgct gtcggtggtc ggggtctcgc
     7141 gctcctcctt ggcgacctgg tgctcctggt cctccaacac gcagcccagg gtgaagtggc
     7201 ccacggccga cagggcgtac aacgcgttct ccagcgagaa gccctgctgg cacaggaagg
     7261 ccaactggtt ctccagggtc tcgtactgct tctcggtcgg ccgggtgccc aggtgcacct
     7321 tggcgccgtc gcggtggctc agcaaggcgc agcggaagct cttggcgttg ttgcgcagga
     7381 agtcctgcca gctctcgccc tccagcgggc agaagtgggt gtggtggcgg tccaacatct
     7441 cgatggccag ggcgtcgagc agggcccgct tgttcttgac gtgccagtac agggtcggct
     7501 gctcgacgcc cagcttctgg gccagcttgc gggtggtcag gccctcgatg ccgacctcgt
     7561 tcagcagctc cagggcggag ttgatgacct tggacttgtc caggcgggac attgcgaagt
     7621 gattcctcct gatggtaccg gatccgtggc aggagggtcg cctgcgtcgg caactcgccc
     7681 aggctacacg agagccctcg actgcgaaaa tgccgaccag gcaccaccgc actccaggct
     7741 taacccggtg cgacgatcca atattacgag atctgacaag tttgtacaaa aaagcaggct
     7801 accaggccta gatctgggga ccctagaggt cccctt
//
