LOCUS       sequence-211575-        9122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Analogous to pMINTNH3 but with a C-terminal 3C cleavage site and a 
            GFP-His10 tag fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9122)
  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 9122)
  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: pMINTC3GH (Hyg)
FEATURES             Location/Qualifiers
     source          1..9122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(298..322)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(401..706)
                     /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(503..522)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1048..1707)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1622..1641
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1761..1767,1768..1785,1786..1791))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1816..1940)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1959..1979)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1983..2003)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2030..2053
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2060..2770
                     /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(2087..2115)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2710..2730
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2777..2806
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     rep_origin      complement(3076..3664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3156..3175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5668..5687
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(7623..7669)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7925..7947
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7944..7960
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7944..7960
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    7980..8000
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     8053..8069
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8054..8070
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9007..9031
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtcggcaccg tcacggccgt gggaggcggc acgatccgcg acgtgatgat cggccgcatc
       61 cccacggtgc tgcgcagtga gctctacgcc atcccggcgt tgatctgtgc gttcgcacgc
      121 acaggcccgg tgtgagaagg gtctctgacg agcgggagaa ctccctatca gtgatagagt
      181 ttgtcctccc tatcagtgat agataggctc tgggagtacc cgtgtgtacg accagcacgg
      241 catacatcat ttcgacgccg agagattcgc cgcccgaaat gagcacgatc cgcatgcacc
      301 cagctttctt gtacaaagtg ggctcgaggc atgcgaagga gatatacata tgagttgaag
      361 agcgacctgc agactggctg tgtataaggg agcctgacat ttatattccc cagaacatca
      421 ggttaatggc gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga
      481 taacggagac cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga
      541 tatgcaccac cgggtaaagt tcacgggaga ctttatctga cagcagacgt gcactggcca
      601 gggggatcac catccgtcgc ccgggcgtgt caataatatc actctgtaca tccacaaaca
      661 gacgataacg gctctctctt ttataggtgt aaaccttaaa ctgcatttca ccagcccctg
      721 ttctcgtcag caaaagagcc gttcatttca ataaaccggg cgacctcagc catcccttcc
      781 tgattttccg ctttccagcg ttcggcacgc agacgacggg cttcattctg catggttgtg
      841 cttaccagac cggagatatt gacatcatat atgccttgag caactgatag ctgtcgctgt
      901 caactgtcac tgtaatacgc tgcttcatag catacctctt tttgacatac ttcgggtata
      961 catatcagta tatattctta taccgcaaaa atcagcgcgc aaatacgcat actgttatct
     1021 ggcttttagt aagccggatc cacgcgttta cgccccgccc tgccactcat cgcagtactg
     1081 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     1141 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     1201 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     1261 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     1321 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     1381 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     1441 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     1501 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     1561 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     1621 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     1681 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgacgg
     1741 atcctaactc aaaatccaca cattatacga gccggaagca taaagtgtaa agcctggggt
     1801 gcctaatgcg gccgccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt
     1861 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
     1921 gcttttttgt acaaacttgt gatatcgaat tcgctagcta cctttctctt cttttttgga
     1981 tctacctttc tcttcttttt tggggatcca ctagttcagc tcttctgcat tagaagtttt
     2041 gtttcaaggt ccacaattca gcaaaggaga agaacttttc actggagttg tcccaattct
     2101 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
     2161 tgatgctaca tacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt
     2221 tccttggcca acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc
     2281 ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     2341 acgcactata tctttcaaag atgacgggaa ctacaagacg cgtgctgaag tcaagtttga
     2401 aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag aagatggaaa
     2461 cattctcgga cacaaactag agtacaacta taactcacac aatgtataca tcacggcaga
     2521 caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacattg aagatggttc
     2581 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     2641 accagacaac cattacctgt cgacacaatc tgccctttcg aaagatccca acgaaaagcg
     2701 tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga
     2761 gctctacaaa actagtcatc atcatcacca tcatcatcac catcattaat tatacatagt
     2821 tgcttaagct gcaggggacc ctagaggtcc ccttttttat tttaaaaatt ttttataacc
     2881 ggtaagaagt ccaacacgcc gctcctcccc gatgttgcgt agccgcttca agcttaagca
     2941 acggccccag tgagtagcga ctttggatcc ggtacccgca acggtcaacc agcgtctacg
     3001 cgtttcactg gccgacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3061 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3121 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3181 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3241 tctcccctcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3301 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3361 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3421 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3481 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3541 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3601 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3661 tcaagaagat cctttgatct tttctacggg gtctgacgct catggaacta gtgaggtctg
     3721 cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag
     3781 aaagtgaggg agccacggtt ggtgagagct ttgttgtagg tggaccagtt ggtgattttg
     3841 aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc
     3901 aactcagcaa aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc
     3961 tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat
     4021 gaaactgcaa gtattctaga ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4261 cgacgtggcc gaccagcccg tcatcgtcaa cgcctgatcc gcggtgcgga caggccgtgt
     4321 cgcgaccggc cgtgcggaat taagccggcc cgtaccctgt gaatagaggt ccgctgtgac
     4381 acaagaatct ctgttacttc tcgaccgtat tgattcggat gattcctacg cgagcctgcg
     4441 gaacgaccag gaattctggg agccgctggc ccgccgagcc ctggaggagc tcgggctgcc
     4501 ggtgccgccg gtgctgcggg tgcccggcga gagcaccaac cccgtactgg tcggcgagcc
     4561 cggcccggtg atcaagctgt tcggcgagca ctggtgcggt ccggagagcc tcgcgtcgga
     4621 gtcggaggcg tacgcggtcc tggcggacgc cccggtgccg gtgccccgcc tcctcggccg
     4681 cggcgagctg cggcccggca ccggagcctg gccgtggccc tacctggtga tgagccggat
     4741 gaccggcacc acctggcggt ccgcgatgga cggcacgacc gaccggaacg cgctgctcgc
     4801 cctggcccgc gaactcggcc gggtgctcgg ccggctgcac agggtgccgc tgaccgggaa
     4861 caccgtgctc accccccatt ccgaggtctt cccggaactg ctgcgggaac gccgcgcggc
     4921 gaccgtcgag gaccaccgcg ggtggggcta cctctcgccc cggctgctgg accgcctgga
     4981 ggactggctg ccggacgtgg acacgctgct ggccggccgc gaaccccggt tcgtccacgg
     5041 cgacctgcac gggaccaaca tcttcgtgga cctggccgcg accgaggtca ccgggatcgt
     5101 cgacttcacc gacgtctatg cgggagactc ccgctacagc ctggtgcaac tgcatctcaa
     5161 cgccttccgg ggcgaccgcg agatcctggc cgcgctgctc gacggggcgc agtggaagcg
     5221 gaccgaggac ttcgcccgcg aactgctcgc cttcaccttc ctgcacgact tcgaggtgtt
     5281 cgaggagacc ccgctggatc tctccggctt caccgatccg gaggaactgg cgcagttcct
     5341 ctgggggccg ccggacaccg cccccggcgc ctgacgcccc gggcccgcgc cgaagcccgc
     5401 tgctgcgagc ccggagcggg ccggccgacg gcggtgcggg cccggcggcg gacgctcagc
     5461 agcggcgggc gtgaaaggcc ctggcatcct cgatcatctc ctccagggtg ggtcggccgg
     5521 gcttccatcc cagctcggca aggatcaacg aggacagtcg cacgacgaag ttcttctgga
     5581 tcgcgcccgt gctggaagca ctcaacctcg aagcgtgtgg ttgcggagcc atctagcaac
     5641 cacacgacca tggtactccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     5701 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     5761 gagattttga gacacaacgt ggctttcccc ccccccgacg tcaggtggct agctggcggg
     5821 aacggcaatg ctcctggcgg cggcggggct ggtgcccagg tctccgcaca gagcggcggt
     5881 gctggcgctc gcggccaggc gtggttcttc gcgtactgac aagaaacccc cctctttagg
     5941 actcagtgtc cttgggaggg gggctttttg cgtttcagga ggtcttggcc agcttggaca
     6001 tcgcctcagc gatagcctcg tcgcgggcct cagacgccat ctggtacttc atcgccatcc
     6061 taggagtcgt gtgaccgaga cgggccatca gctccttggt cgtcgcacct gcctgagcgg
     6121 cgaacgtagc gccgacagcg cggaggtcgt ggatgcggag ttccggccga ccgatcttgg
     6181 cgtagccacg cttcagcgac ttggtgaacg cggacttcga cagccggttg ccctgcgtcg
     6241 tggtcaccag gaatgcctcg gggcccttgt tcatcttcgt acggtccttc atgtgcgctc
     6301 ggatcatctc cgcgacgtga ggcggaaccg tcacaggacg cttcgaccgg acggtcttgg
     6361 cgttgccaac gacgatcttg ttccccacgc gggaagcgcc acggcgcacc cggagcttca
     6421 tcgtcatgcc gtcgtccacg atgtccttgc ggcgaagctc gatcagctct ccgaaccgga
     6481 ggctcgtcca cgccaggatg tatgccgcga tccggtagtg cacgaagatc tcagcggcga
     6541 cgatgtccag ctcctcaggc gtcagcgcct ctacgtcgcg ctcatcggct gccttctgct
     6601 cgatccggca cgggttctct gcgatcagct tgtcctcgac cgctgtgttc atcaccgccc
     6661 ggaggacgtt gtaggcatgc cggcgggcag tcgggtgctt cctacccatc ccggcccacc
     6721 acgcacgcac cagagctggc gtcatctctg tgaccgccac ttcacctagc accgggtaga
     6781 tgcggcgctc cgcgtgcccg ctgtacagat ccctggtgcc gtctgcgagg tcgcgctcca
     6841 cgagccactt ccgggtgtac tcctccagcg tgatggcgct ggcggctgcc ttcttcgccc
     6901 ggtcctgtgg aggggtccag gtctccatct cgatgagccg cttctcgccc gcgagccagg
     6961 cttcggcgtc catcttgttg tcgtaggtct gcagcgcgta gtacctcaca ccgtcctgcg
     7021 ggttgacgta tgaggcttgg atcctcccgc tgcgctgagt cttcagcgat ccccatccgc
     7081 gacgtgccaa ctaggtctcc tctcgtcgtg aacaaggcta ccgggttgca actcctgtgc
     7141 aactctcagg cttcaacgcg cttctacgac ctgcaatttc tttccactta gaggatgcag
     7201 ccgagagggg gtaaaaacct atcttgaccg gcccatatat gtggtcggca gacacccatt
     7261 cttccaaact agctacgcgg gttcgattcc cgtcgcccgc tccgctggtc agagggtgtt
     7321 ttcgccctct ggccattttt ctttccaggg gtctgcaact cttgtgcgac tcttctgacc
     7381 tgggcatacg cggttgcaac gcatccctga tctggctact ttcgatgctg acaaacgaat
     7441 agagcccccc gcctgcgcga acagacgagg ggcattcaca ccagattgga gctggtgcag
     7501 tgaagagaat agaccgggac aaggttgcac cgggagttgc agcggtcgga accctcgccg
     7561 tcggcgggct ggcgttcgcc ctgtcgttca cggctctagc tgatcaccgc ggccatgatg
     7621 gcataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag
     7681 tcgatcgtac gctagttaac tacgtcgaca tcgatttaat taatatgcat caattgattt
     7741 aaatactagt gttaactatt taattgggga ccctagaggt cccctttttt attttaaaaa
     7801 ttttttcaag gggacctcta gggtccccag ctggctagcg agtcatgagg tcaccatgtt
     7861 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7921 taccagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctatc
     7981 aactttgtat agaaaagttg catgaccaac ttcgataacg ttctcggctc gatgatcccc
     8041 gggggccccc cctcgaggtc gacggtatcg atttaattaa tatgcatcaa ttgatttaaa
     8101 tactagtgtt aactatttaa ttggggaccc tagaggtccc cttttttatt ttaaaaattt
     8161 tttcaagggg acctctaggg tccccagctg gctagcgagt catgaggtca ccatggtcat
     8221 taggagccgc tctcgcactt cagctgcttc tccaagccgc agatgatcag ctccaggccg
     8281 aacaagaagg ccggctcggc gccctggtgg tcgaacagct cgatggcctg gcgcagcaac
     8341 ggcggcatgc tgtcggtggt cggggtctcg cgctcctcct tggcgacctg gtgctcctgg
     8401 tcctccaaca cgcagcccag ggtgaagtgg cccacggccg acagggcgta caacgcgttc
     8461 tccagcgaga agccctgctg gcacaggaag gccaactggt tctccagggt ctcgtactgc
     8521 ttctcggtcg gccgggtgcc caggtgcacc ttggcgccgt cgcggtggct cagcaaggcg
     8581 cagcggaagc tcttggcgtt gttgcgcagg aagtcctgcc agctctcgcc ctccagcggg
     8641 cagaagtggg tgtggtggcg gtccaacatc tcgatggcca gggcgtcgag cagggcccgc
     8701 ttgttcttga cgtgccagta cagggtcggc tgctcgacgc ccagcttctg ggccagcttg
     8761 cgggtggtca ggccctcgat gccgacctcg ttcagcagct ccagggcgga gttgatgacc
     8821 ttggacttgt ccaggcggga cattgcgaag tgattcctcc tgatggtacc ggatccgtgg
     8881 caggagggtc gcctgcgtcg gcaactcgcc caggctacac gagagccctc gactgcgaaa
     8941 atgccgacca ggcaccaccg cactccaggc ttaacccggt gcgacgatcc aatattacga
     9001 gatctgacaa gtttgtacaa aaaagcaggc taccaggcct agatctgggg accctagagg
     9061 tccccttttt tattttaaaa attttttcac gtgccagccg attggactag aaatattgga
     9121 tc
//
