LOCUS       sequence-211291-        7533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pMINTNH3 but with a C-terminal 3C cleavage site and 
            His10 tag. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7533)
  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 7533)
  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: pMINTC3H
FEATURES             Location/Qualifiers
     source          1..7533
                     /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(6089..6135)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6391..6413
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6410..6426
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6410..6426
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6446..6466
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     6519..6535
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6520..6536
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7473..7497
                     /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 ttggccagct tggacatcgc ctcagcgata gcctcgtcgc gggcctcaga
     4501 cgccatctgg tacttcatcg ccatcctagg agtcgtgtga ccgagacggg ccatcagctc
     4561 cttggtcgtc gcacctgcct gagcggcgaa cgtagcgccg acagcgcgga ggtcgtggat
     4621 gcggagttcc ggccgaccga tcttggcgta gccacgcttc agcgacttgg tgaacgcgga
     4681 cttcgacagc cggttgccct gcgtcgtggt caccaggaat gcctcggggc ccttgttcat
     4741 cttcgtacgg tccttcatgt gcgctcggat catctccgcg acgtgaggcg gaaccgtcac
     4801 aggacgcttc gaccggacgg tcttggcgtt gccaacgacg atcttgttcc ccacgcggga
     4861 agcgccacgg cgcacccgga gcttcatcgt catgccgtcg tccacgatgt ccttgcggcg
     4921 aagctcgatc agctctccga accggaggct cgtccacgcc aggatgtatg ccgcgatccg
     4981 gtagtgcacg aagatctcag cggcgacgat gtccagctcc tcaggcgtca gcgcctctac
     5041 gtcgcgctca tcggctgcct tctgctcgat ccggcacggg ttctctgcga tcagcttgtc
     5101 ctcgaccgct gtgttcatca ccgcccggag gacgttgtag gcatgccggc gggcagtcgg
     5161 gtgcttccta cccatcccgg cccaccacgc acgcaccaga gctggcgtca tctctgtgac
     5221 cgccacttca cctagcaccg ggtagatgcg gcgctccgcg tgcccgctgt acagatccct
     5281 ggtgccgtct gcgaggtcgc gctccacgag ccacttccgg gtgtactcct ccagcgtgat
     5341 ggcgctggcg gctgccttct tcgcccggtc ctgtggaggg gtccaggtct ccatctcgat
     5401 gagccgcttc tcgcccgcga gccaggcttc ggcgtccatc ttgttgtcgt aggtctgcag
     5461 cgcgtagtac ctcacaccgt cctgcgggtt gacgtatgag gcttggatcc tcccgctgcg
     5521 ctgagtcttc agcgatcccc atccgcgacg tgccaactag gtctcctctc gtcgtgaaca
     5581 aggctaccgg gttgcaactc ctgtgcaact ctcaggcttc aacgcgcttc tacgacctgc
     5641 aatttctttc cacttagagg atgcagccga gagggggtaa aaacctatct tgaccggccc
     5701 atatatgtgg tcggcagaca cccattcttc caaactagct acgcgggttc gattcccgtc
     5761 gcccgctccg ctggtcagag ggtgttttcg ccctctggcc atttttcttt ccaggggtct
     5821 gcaactcttg tgcgactctt ctgacctggg catacgcggt tgcaacgcat ccctgatctg
     5881 gctactttcg atgctgacaa acgaatagag ccccccgcct gcgcgaacag acgaggggca
     5941 ttcacaccag attggagctg gtgcagtgaa gagaatagac cgggacaagg ttgcaccggg
     6001 agttgcagcg gtcggaaccc tcgccgtcgg cgggctggcg ttcgccctgt cgttcacggc
     6061 tctagctgat caccgcggcc atgatggcat aaaacgaaag gcccagtctt tcgactgagc
     6121 ctttcgtttt atttgatgcc tggcagtcga tcgtacgcta gttaactacg tcgacatcga
     6181 tttaattaat atgcatcaat tgatttaaat actagtgtta actatttaat tggggaccct
     6241 agaggtcccc ttttttattt taaaaatttt ttcaagggga cctctagggt ccccagctgg
     6301 ctagcgagtc atgaggtcac catgttcttt cctgcgttat cccctgattc tgtggataac
     6361 cgtattaccg cctttgagtg agctgatacc agcggataac aatttcacac aggaaacagc
     6421 tatgaccatg attacgccaa gctatcaact ttgtatagaa aagttgcatg accaacttcg
     6481 ataacgttct cggctcgatg atccccgggg gccccccctc gaggtcgacg gtatcgattt
     6541 aattaatatg catcaattga tttaaatact agtgttaact atttaattgg ggaccctaga
     6601 ggtccccttt tttattttaa aaattttttc aaggggacct ctagggtccc cagctggcta
     6661 gcgagtcatg aggtcaccat ggtcattagg agccgctctc gcacttcagc tgcttctcca
     6721 agccgcagat gatcagctcc aggccgaaca agaaggccgg ctcggcgccc tggtggtcga
     6781 acagctcgat ggcctggcgc agcaacggcg gcatgctgtc ggtggtcggg gtctcgcgct
     6841 cctccttggc gacctggtgc tcctggtcct ccaacacgca gcccagggtg aagtggccca
     6901 cggccgacag ggcgtacaac gcgttctcca gcgagaagcc ctgctggcac aggaaggcca
     6961 actggttctc cagggtctcg tactgcttct cggtcggccg ggtgcccagg tgcaccttgg
     7021 cgccgtcgcg gtggctcagc aaggcgcagc ggaagctctt ggcgttgttg cgcaggaagt
     7081 cctgccagct ctcgccctcc agcgggcaga agtgggtgtg gtggcggtcc aacatctcga
     7141 tggccagggc gtcgagcagg gcccgcttgt tcttgacgtg ccagtacagg gtcggctgct
     7201 cgacgcccag cttctgggcc agcttgcggg tggtcaggcc ctcgatgccg acctcgttca
     7261 gcagctccag ggcggagttg atgaccttgg acttgtccag gcgggacatt gcgaagtgat
     7321 tcctcctgat ggtaccggat ccgtggcagg agggtcgcct gcgtcggcaa ctcgcccagg
     7381 ctacacgaga gccctcgact gcgaaaatgc cgaccaggca ccaccgcact ccaggcttaa
     7441 cccggtgcga cgatccaata ttacgagatc tgacaagttt gtacaaaaaa gcaggctacc
     7501 aggcctagat ctggggaccc tagaggtccc ctt
//
