LOCUS       sequence-264216-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative vector for expression in mycobacteria based on the Tet 
            promoter. Contains an N-terminal His10 Tag followed by a 3C cleavage
            site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7537)
  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 7537)
  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: pMINTNH3
FEATURES             Location/Qualifiers
     source          1..7537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(597..616)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             608..1423
                     /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     complement(681..700)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1754..2342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2243..2262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2641..2661
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2665..2685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    2704..2828
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2853..2858,2859..2876,2877..2883)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2937..3596
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3003..3022)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3938..4243
                     /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     4122..4141
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4292..4315)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(4316..4345)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     protein_bind    4376..4400
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4789..4813)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5750..5766)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5751..5767)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(5820..5840)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(5860..5876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5860..5876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5873..5895)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     terminator      6151..6197
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtacgaagat gaacaagggc cccgaggcat tcctggtgac cacgacgcag ggcaaccggc
       61 tgtcgaagtc cgcgttcacc aagtcgctga agcgtggcta cgccaagatc ggtcggccgg
      121 aactccgcat ccacgacctc cgcgctgtcg gcgctacgtt cgccgctcag gcaggtgcga
      181 cgaccaagga gctgatggcc cgtctcggtc acacgactcc taggatggcg atgaagtacc
      241 agatggcgtc tgaggcccgc gacgaggcta tcgctgaggc gatgtccaag ctggccaaga
      301 cctcctgaaa cgcaaaaagc ccccctccca aggacactga gtcctaaaga ggggggtttc
      361 ttgtcagtac gcgaagaacc acgcctggcc gcgagcgcca gcaccgccgc tctgtgcgga
      421 gacctgggca ccagccccgc cgccgccagg agcattgccg ttcccgccag ctagccacct
      481 gacgtcgggg gggggggaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
      541 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
      601 gggtgttatg agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa
      661 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
      721 gacaatctat cgcttgtatg ggaagcccca tgcgccagag ttgtttctga aacatggcaa
      781 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
      841 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
      901 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
      961 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1021 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1081 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1141 ctggaaagaa atgcataatc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     1201 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     1261 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     1321 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     1381 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     1441 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaataaat
     1501 cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc
     1561 gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcaccaac
     1621 cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta tgagtcagca
     1681 acaccttctt cacgaggcag acctcactag ttccatgagc gtcagacccc gtagaaaaga
     1741 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1801 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1861 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     1921 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1981 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2041 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2101 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2161 cgcttcccga ggggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2221 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2281 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2341 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2401 tgtcggccag tgaaacgcgt agacgctggt tgaccgttgc gggtaccgga tccaaagtcg
     2461 ctactcactg gggccgttgc ttaagcttga agcggctacg caacatcggg gaggagcggc
     2521 gtgttggact tcttaccggt tataaaaaat ttttaaaata aaaaagggga cctctagggt
     2581 cccctgcagc ttaagcaact atgtataatt attatgcaga agagctgaac tagtggatcc
     2641 ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa aggtagctag cgaattcgat
     2701 atcacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata
     2761 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     2821 tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata
     2881 atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg
     2941 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     3001 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     3061 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     3121 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
     3181 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     3241 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     3301 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     3361 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     3421 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     3481 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     3541 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaacgc
     3601 gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     3661 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
     3721 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     3781 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     3841 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     3901 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     3961 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     4021 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     4081 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     4141 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     4201 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     4261 acacagccag tctgcaggtc gctcttcaac ttggaccttg aaacaaaact tctaaatgat
     4321 gatgatgatg gtgatgatga tggtgcatat gtatatctcc ttcgcatgcc tcgagcccac
     4381 tttgtacaag aaagctgggt gcatgcggat cgtgctcatt tcgggcggcg aatctctcgg
     4441 cgtcgaaatg atgtatgccg tgctggtcgt acacacgggt actcccagag cctatctatc
     4501 actgataggg aggacaaact ctatcactga tagggagttc tcccgctcgt cagagaccct
     4561 tctcacaccg ggcctgtgcg tgcgaacgca cagatcaacg ccgggatggc gtagagctca
     4621 ctgcgcagca ccgtggggat gcggccgatc atcacgtcgc ggatcgtgcc gcctcccacg
     4681 gccgtgacgg tgccgacgat ccaatatttc tagtccaatc ggctggcacg tgaaaaaatt
     4741 tttaaaataa aaaaggggac ctctagggtc cccagatcta ggcctggtag cctgcttttt
     4801 tgtacaaact tgtcagatct cgtaatattg gatcgtcgca ccgggttaag cctggagtgc
     4861 ggtggtgcct ggtcggcatt ttcgcagtcg agggctctcg tgtagcctgg gcgagttgcc
     4921 gacgcaggcg accctcctgc cacggatccg gtaccatcag gaggaatcac ttcgcaatgt
     4981 cccgcctgga caagtccaag gtcatcaact ccgccctgga gctgctgaac gaggtcggca
     5041 tcgagggcct gaccacccgc aagctggccc agaagctggg cgtcgagcag ccgaccctgt
     5101 actggcacgt caagaacaag cgggccctgc tcgacgccct ggccatcgag atgttggacc
     5161 gccaccacac ccacttctgc ccgctggagg gcgagagctg gcaggacttc ctgcgcaaca
     5221 acgccaagag cttccgctgc gccttgctga gccaccgcga cggcgccaag gtgcacctgg
     5281 gcacccggcc gaccgagaag cagtacgaga ccctggagaa ccagttggcc ttcctgtgcc
     5341 agcagggctt ctcgctggag aacgcgttgt acgccctgtc ggccgtgggc cacttcaccc
     5401 tgggctgcgt gttggaggac caggagcacc aggtcgccaa ggaggagcgc gagaccccga
     5461 ccaccgacag catgccgccg ttgctgcgcc aggccatcga gctgttcgac caccagggcg
     5521 ccgagccggc cttcttgttc ggcctggagc tgatcatctg cggcttggag aagcagctga
     5581 agtgcgagag cggctcctaa tgaccatggt gacctcatga ctcgctagcc agctggggac
     5641 cctagaggtc cccttgaaaa aatttttaaa ataaaaaagg ggacctctag ggtccccaat
     5701 taaatagtta acactagtat ttaaatcaat tgatgcatat taattaaatc gataccgtcg
     5761 acctcgaggg ggggcccccg gggatcatcg agccgagaac gttatcgaag ttggtcatgc
     5821 aacttttcta tacaaagttg atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5881 aaattgttat ccgctggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5941 aggggataac gcaggaaaga acatggtgac ctcatgactc gctagccagc tggggaccct
     6001 agaggtcccc ttgaaaaaat ttttaaaata aaaaagggga cctctagggt ccccaattaa
     6061 atagttaaca ctagtattta aatcaattga tgcatattaa ttaaatcgat gtcgacgtag
     6121 ttaactagcg tacgatcgac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     6181 tgggcctttc gttttatgcc atcatggccg cggtgatcag ctagagccgt gaacgacagg
     6241 gcgaacgcca gcccgccgac ggcgagggtt ccgaccgctg caactcccgg tgcaaccttg
     6301 tcccggtcta ttctcttcac tgcaccagct ccaatctggt gtgaatgccc ctcgtctgtt
     6361 cgcgcaggcg gggggctcta ttcgtttgtc agcatcgaaa gtagccagat cagggatgcg
     6421 ttgcaaccgc gtatgcccag gtcagaagag tcgcacaaga gttgcagacc cctggaaaga
     6481 aaaatggcca gagggcgaaa acaccctctg accagcggag cgggcgacgg gaatcgaacc
     6541 cgcgtagcta gtttggaaga atgggtgtct gccgaccaca tatatgggcc ggtcaagata
     6601 ggtttttacc ccctctcggc tgcatcctct aagtggaaag aaattgcagg tcgtagaagc
     6661 gcgttgaagc ctgagagttg cacaggagtt gcaacccggt agccttgttc acgacgagag
     6721 gagacctagt tggcacgtcg cggatgggga tcgctgaaga ctcagcgcag cgggaggatc
     6781 caagcctcat acgtcaaccc gcaggacggt gtgaggtact acgcgctgca gacctacgac
     6841 aacaagatgg acgccgaagc ctggctcgcg ggcgagaagc ggctcatcga gatggagacc
     6901 tggacccctc cacaggaccg ggcgaagaag gcagccgcca gcgccatcac gctggaggag
     6961 tacacccgga agtggctcgt ggagcgcgac ctcgcagacg gcaccaggga tctgtacagc
     7021 gggcacgcgg agcgccgcat ctacccggtg ctaggtgaag tggcggtcac agagatgacg
     7081 ccagctctgg tgcgtgcgtg gtgggccggg atgggtagga agcacccgac tgcccgccgg
     7141 catgcctaca acgtcctccg ggcggtgatg aacacagcgg tcgaggacaa gctgatcgca
     7201 gagaacccgt gccggatcga gcagaaggca gccgatgagc gcgacgtaga ggcgctgacg
     7261 cctgaggagc tggacatcgt cgccgctgag atcttcgtgc actaccggat cgcggcatac
     7321 atcctggcgt ggacgagcct ccggttcgga gagctgatcg agcttcgccg caaggacatc
     7381 gtggacgacg gcatgacgat gaagctccgg gtgcgccgtg gcgcttcccg cgtggggaac
     7441 aagatcgtcg ttggcaacgc caagaccgtc cggtcgaagc gtcctgtgac ggttccgcct
     7501 cacgtcgcgg agatgatccg agcgcacatg aaggacc
//
