LOCUS       sequence-211300-        7840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pMINTNH3, but containing oriM for episomal replication 
            in mycobacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7840)
  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 7840)
  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: pMEXNH3
FEATURES             Location/Qualifiers
     source          1..7840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(328..352)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             383..412
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             413..436
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(485..790)
                     /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(587..606)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1132..1791)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1706..1725
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1845..1851,1852..1869,1870..1875))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1900..2024)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2043..2063)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2067..2087)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(2386..2974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2466..2485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3305..4120)
                     /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     4028..4047
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4112..4131
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(6371..6417)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6673..6695
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6692..6708
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6692..6708
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6728..6748
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     6801..6817
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6802..6818
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7755..7779
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gccagccgat tggactagaa atattggatc gtcggcaccg tcacggccgt gggaggcggc
       61 acgatccgcg acgtgatgat cggccgcatc cccacggtgc tgcgcagtga gctctacgcc
      121 atcccggcgt tgatctgtgc gttcgcacgc acaggcccgg tgtgagaagg gtctctgacg
      181 agcgggagaa ctccctatca gtgatagagt ttgtcctccc tatcagtgat agataggctc
      241 tgggagtacc cgtgtgtacg accagcacgg catacatcat ttcgacgccg agagattcgc
      301 cgcccgaaat gagcacgatc cgcatgcacc cagctttctt gtacaaagtg ggctcgaggc
      361 atgcgaagga gatatacata tgcaccatca tcatcaccat catcatcatc atttagaagt
      421 tttgtttcaa ggtccaagtt gaagagcgac ctgcagactg gctgtgtata agggagcctg
      481 acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc
      541 tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata tcggtggtca
      601 tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg gagactttat
      661 ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc gtgtcaataa
      721 tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag gtgtaaacct
      781 taaactgcat ttcaccagcc cctgttctcg tcagcaaaag agccgttcat ttcaataaac
      841 cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc acgcagacga
      901 cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc atatatgcct
      961 tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc atagcatacc
     1021 tctttttgac atacttcggg tatacatatc agtatatatt cttataccgc aaaaatcagc
     1081 gcgcaaatac gcatactgtt atctggcttt tagtaagccg gatccacgcg tttacgcccc
     1141 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     1201 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     1261 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     1321 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     1381 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     1441 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     1501 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     1561 cattgccata cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     1621 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     1681 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     1741 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     1801 cttagctcct gaaaatctcg acggatccta actcaaaatc cacacattat acgagccgga
     1861 agcataaagt gtaaagcctg gggtgcctaa tgcggccgcc atagtgactg gatatgttgt
     1921 gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt
     1981 atatcatttt acgtttctcg ttcagctttt ttgtacaaac ttgtgatatc gaattcgcta
     2041 gctacctttc tcttcttttt tggatctacc tttctcttct tttttgggga tccactagtt
     2101 cagctcttct gcataataat tatacatagt tgcttaagct gcaggggacc ctagaggtcc
     2161 ccttttttat tttaaaaatt ttttataacc ggtaagaagt ccaacacgcc gctcctcccc
     2221 gatgttgcgt agccgcttca agcttaagca acggccccag tgagtagcga ctttggatcc
     2281 ggtacccgca acggtcaacc agcgtctacg cgtttcactg gccgacatgt gagcaaaagg
     2341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2521 cctgccgctt accggatacc tgtccgcctt tctcccctcg ggaagcgtgg cgctttctca
     2581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2821 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3001 gtctgacgct catggaacta gtgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     3061 caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt ggtgagagct
     3121 ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg
     3181 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     3241 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     3301 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3361 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3421 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3481 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3541 cgactgaatc cggtgagaat ggcaaaagat tatgcatttc tttccagact tgttcaacag
     3601 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3661 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3721 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3781 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3841 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3901 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3961 gaaacaactc tggcgcatgg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     4021 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4081 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4141 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     4201 atcagagatt ttgagacaca acgtggcttt cccccccccc cgacgtcagg tggctagctg
     4261 gcgggaacgg caatgctcct ggcggcggcg gggctggtgc ccaggtctcc gcacagagcg
     4321 gcggtgctgg cgctcgcggc caggcgtggt tcttcgcgta ctgacaagaa acccccctct
     4381 ttaggactca gtgtccttgg gaggggggct ttttgcgttt caggaggtct tggccaggca
     4441 tgccacggat gccaccacag gcactacaac ggagttcgcc acgtacatca ccacaaccac
     4501 cgattctggc ggtgagctcc ccgatattca gcggaaatgg cttggtatcg accaagattc
     4561 gtagaacccc gtctcgtctg gctggtattc aaaacgggcg caacgaaaca cgcaacgaga
     4621 caggcatggc ccaaaccaga aaactagcgt ctaccaggac ttttacctgt ccgacccgtt
     4681 gcaacggaac cccccacgga acccccgcga cacccgctcc ccaattgcgt tagaacagcg
     4741 gtggattgtc ggcttcgttg tgggcctttt gagccgcttc ctgttctgcc gcacgctctt
     4801 tcctcgcccg atagccgagt cgcttaacgg tgtccagatg cagcccgaaa tgtttggccg
     4861 tttgcggcca agagtggccc tcgtcgtcgt gataggcgcg gatgcgttcg cggcgtgcag
     4921 cctgctcggc gagccactcg ctgcgttcct gcgccacgag ccggacgacg tggcgttcgg
     4981 atagtccggt gattcgagcg ccttcggcgg cggtcacgcg ccgctttttg cggacagtcg
     5041 gctgccggtt gtagccgtcg ctgtagccgt cgctcatagc aatgcctcca tgcatggctg
     5101 acgcggactt tgcgcgccgc gcaactgtgc tcgccgccgt gcgcgctgct gcgcccttcc
     5161 gcgagatggc cgactggcgc gcactgagtg tggcctcgta gaccacgatc ccgtccgccc
     5221 aaatgcgcga cttggttgtg atccaacgcc aaatgctgtt ggcgatggcg cggacctcgc
     5281 tgtccggtag cggtccggga cacacgtcgt tgcacggaaa ttcggcgttt cgcgcgtggc
     5341 actcggcata gatcgcgcgg ccgagtccgt ccacgttccg ggtcggcagg tagatccgca
     5401 tgagggcggg acgataggcc cacaacctga cggaatcgaa cagtgcgcaa ttccgcccta
     5461 gcggcgtcgg agccgctttg tacgtggtct gctgacgcca gcgcggcggt ggcatgttcg
     5521 cgccgagctc ggcctcgatg tggctgagtg tgtagagatc tgagtggagc cattccgttt
     5581 cccaggcgat gtggccgggg tttttggtca tgaggcctga gtaactgcgg tcgccatcga
     5641 cggcgcgccg aaggccttcg gcgcacgccg ccatgtatgc gagcggctta cgccgcgcgt
     5701 attcggtgcg tggaacaggg gcgttgagtg cccacactgc gtgtgcgtgg ccgttggcgc
     5761 gattgcccac gatcgcgttg ggcagcggat gggacccccg ggcgctgagc gctcggagcg
     5821 ctgcgtctgg atggtctacg tccacgacca gcaggtttgc cagcgctgtt gggttcgcct
     5881 cgatgtaccg gcggcctagg gccgacgcgc ggctttggcg gtagatcccc tcgagcagat
     5941 cgtcgcttgc cagcggccag tacggcagcc agagctgctc aaattcgtcg gcgacgtggc
     6001 tcacgcttgg tagtagacca cgattaatca ccggtgtatg gtccgacacg agctccaagt
     6061 cagatatttc gctgaggggc caccccacaa ctgcacactc ccccgctctc ccgtcgagcc
     6121 ctggtggtgg aacaccagcg acagccgagc acccccaacc acctgtacca accaggagga
     6181 acacatgcgt cgtttcgagg acgtttccgg gccgctgaga gccgctgtgg cggccgtaca
     6241 cgccgcctta gacccgttag accccctgcc gcctgaatgc gcgggtacga gccacacagc
     6301 gcccgaactt acggagctgg tgggctcact ctagatcacg gctctagctg atcaccgcgg
     6361 ccatgatggc ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     6421 cctggcagtc gatcgtacgc tagttaacta cgtcgacatc gatttaatta atatgcatca
     6481 attgatttaa atactagtgt taactattta attggggacc ctagaggtcc ccttttttat
     6541 tttaaaaatt ttttcaaggg gacctctagg gtccccagct ggctagcgag tcatgaggtc
     6601 accatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6661 tgagctgata ccagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6721 aagctatcaa ctttgtatag aaaagttgca tgaccaactt cgataacgtt ctcggctcga
     6781 tgatccccgg gggccccccc tcgaggtcga cggtatcgat ttaattaata tgcatcaatt
     6841 gatttaaata ctagtgttaa ctatttaatt ggggacccta gaggtcccct tttttatttt
     6901 aaaaattttt tcaaggggac ctctagggtc cccagctggc tagcgagtca tgaggtcacc
     6961 atggtcatta ggagccgctc tcgcacttca gctgcttctc caagccgcag atgatcagct
     7021 ccaggccgaa caagaaggcc ggctcggcgc cctggtggtc gaacagctcg atggcctggc
     7081 gcagcaacgg cggcatgctg tcggtggtcg gggtctcgcg ctcctccttg gcgacctggt
     7141 gctcctggtc ctccaacacg cagcccaggg tgaagtggcc cacggccgac agggcgtaca
     7201 acgcgttctc cagcgagaag ccctgctggc acaggaaggc caactggttc tccagggtct
     7261 cgtactgctt ctcggtcggc cgggtgccca ggtgcacctt ggcgccgtcg cggtggctca
     7321 gcaaggcgca gcggaagctc ttggcgttgt tgcgcaggaa gtcctgccag ctctcgccct
     7381 ccagcgggca gaagtgggtg tggtggcggt ccaacatctc gatggccagg gcgtcgagca
     7441 gggcccgctt gttcttgacg tgccagtaca gggtcggctg ctcgacgccc agcttctggg
     7501 ccagcttgcg ggtggtcagg ccctcgatgc cgacctcgtt cagcagctcc agggcggagt
     7561 tgatgacctt ggacttgtcc aggcgggaca ttgcgaagtg attcctcctg atggtaccgg
     7621 atccgtggca ggagggtcgc ctgcgtcggc aactcgccca ggctacacga gagccctcga
     7681 ctgcgaaaat gccgaccagg caccaccgca ctccaggctt aacccggtgc gacgatccaa
     7741 tattacgaga tctgacaagt ttgtacaaaa aagcaggcta ccaggcctag atctggggac
     7801 cctagaggtc ccctttttta ttttaaaaat tttttcacgt
//
