LOCUS       sequence-211189-        8266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replicative, acetamide-based expression vector containing a 
            C-terminal 3C cleavage site and a GPF-fusion with a His10-tag. 
            Suitable for high-level expression levels amenable for large scale 
            purification of proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8266)
  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 8266)
  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: pACEC3GH (Hyg)
FEATURES             Location/Qualifiers
     source          1..8266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(438..743)
                     /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(540..559)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1072..1095
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1102..1812
                     /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(1129..1157)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1752..1772
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1819..1848
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     terminator      1907..1953
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      3326..3914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3815..3834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgctgttcg tcgcgatcgc cgcactcggc tactcgtggc acgcgttcac cgccgaggcc
       61 gacccggcgt tcggggtgat ctggctgctg tgggcagtgc tgtggttcat gctgttcctg
      121 ctgctcggcc tggggcacga cgcactgggg cccgccgtcg ggttcgtcgc ggtggccgaa
      181 ggcgtgatca ccgccgccgt gccggccttc ctgatcgtgt cgggcaactg ggaaaccggc
      241 ccgctccccg ccgcggtcat cgccgtgatc ggttttgccg cagttgttct cgcatacccc
      301 atcgggcgcc gtctcgcagc gccgtcagtc accaaccctc caccggccgc gctcgcggcc
      361 accacccgat aagagaaagg gagtccacca tgagttgaag agcctgcaga ctggctgtgt
      421 ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca
      481 ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc
      541 atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca
      601 cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg
      661 ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta
      721 taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcagcaa aagagccgtt
      781 catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc
      841 ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accagaccgg agatattgac
      901 atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc
      961 ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat attcttatac
     1021 cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc gctcttctgc attagaagtt
     1081 ttgtttcaag gtccacaatt cagcaaagga gaagaacttt tcactggagt tgtcccaatt
     1141 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     1201 ggtgatgcta catacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
     1261 gttccttggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
     1321 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     1381 gaacgcacta tatctttcaa agatgacggg aactacaaga cgcgtgctga agtcaagttt
     1441 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
     1501 aacattctcg gacacaaact agagtacaac tataactcac acaatgtata catcacggca
     1561 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacat tgaagatggt
     1621 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     1681 ttaccagaca accattacct gtcgacacaa tctgcccttt cgaaagatcc caacgaaaag
     1741 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
     1801 gagctctaca aaactagtca tcatcatcac catcatcatc accatcatta ataacagctt
     1861 atcgatgtcg acgtagttaa ctagcgtacg atcgactgcc aggcatcaaa taaaacgaaa
     1921 ggctcagtcg aaagactggg cctttcgttt tatgccatca tggccgcggt gatcagctag
     1981 agggggcgtc aggcgccggg ggcggtgtcc ggcggccccc agaggaactg cgccagttcc
     2041 tccggatcgg tgaagccgga gagatccagc ggggtctcct cgaacacctc gaagtcgtgc
     2101 aggaaggtga aggcgagcag ttcgcgggcg aagtcctcgg tccgcttcca ctgcgccccg
     2161 tcgagcagcg cggccaggat ctcgcggtcg ccccggaagg cgttgagatg cagttgcacc
     2221 aggctgtagc gggagtctcc cgcatagacg tcggtgaagt cgacgatccc ggtgacctcg
     2281 gtcgcggcca ggtccacgaa gatgttggtc ccgtgcaggt cgccgtggac gaaccggggt
     2341 tcgcggccgg ccagcagcgt gtccacgtcc ggcagccagt cctccaggcg gtccagcagc
     2401 cggggcgaga ggtagcccca cccgcggtgg tcctcgacgg tcgccgcgcg gcgttcccgc
     2461 agcagttccg ggaagacctc ggaatggggg gtgagcacgg tgttcccggt cagcggcacc
     2521 ctgtgcagcc ggccgagcac ccggccgagt tcgcgggcca gggcgagcag cgcgttccgg
     2581 tcggtcgtgc cgtccatcgc ggaccgccag gtggtgccgg tcatccggct catcaccagg
     2641 tagggccacg gccaggctcc ggtgccgggc cgcagctcgc cgcggccgag gaggcggggc
     2701 accggcaccg gggcgtccgc caggaccgcg tacgcctccg actccgacgc gaggctctcc
     2761 ggaccgcacc agtgctcgcc gaacagcttg atcaccgggc cgggctcgcc gaccagtacg
     2821 gggttggtgc tctcgccggg cacccgcagc accggcggca ccggcagccc gagctcctcc
     2881 agggctcggc gggccagcgg ctcccagaat tcctggtcgt tccgcaggct cgcgtaggaa
     2941 tcatccgaat caatacggtc gagaagtaac agggattctt gtgtcacagc ggacctctat
     3001 tcacagggta cgggccggct taattccgca cggccggtcg cgacacggcc tgtccgcacc
     3061 gcggatcagg cgttgacgat gacgggctgg tcggccacgt cggggagtca ggcaactatg
     3121 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3181 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3241 aggatctagg tgaagatcct ttttgataat ctcatgctct agttccatga gcgtcagacc
     3301 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3361 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3421 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3481 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3541 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3601 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3661 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3721 gagaaagcgc cacgcttccc gaggggagaa aggcggacag gtatccggta agcggcaggg
     3781 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3841 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3901 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3961 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4021 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcaac gcgtgcggcc
     4081 gcacgcgtga gcccaccagc tccgtaagtt cgggcgctgt gtggctcgta cccgcgcatt
     4141 caggcggcag ggggtctaac gggtctaagg cggcgtgtac ggccgccaca gcggctctca
     4201 gcggcccgga aacgtcctcg aaacgacgca tgtgttcctc ctggttggta caggtggttg
     4261 ggggtgctcg gctgtcgctg gtgttccacc accagggctc gacgggagag cgggggagtg
     4321 tgcagttgtg gggtggcccc tcagcgaaat atctgacttg gagctcgtgt cggaccatac
     4381 accggtgatt aatcgtggtc tactaccaag cgtgagccac gtcgccgacg aatttgagca
     4441 gctctggctg ccgtactggc cgctggcaag cgacgatctg ctcgagggga tctaccgcca
     4501 aagccgcgcg tcggccctag gccgccggta catcgaggcg aacccaacag cgctggcaaa
     4561 cctgctggtc gtggacgtag accatccaga cgcagcgctc cgagcgctca gcgcccgggg
     4621 gtcccatccg ctgcccaacg cgatcgtggg caatcgcgcc aacggccacg cacacgcagt
     4681 gtgggcactc aacgcccctg ttccacgcac cgaatacgcg cggcgtaagc cgctcgcata
     4741 catggcggcg tgcgccgaag gccttcggcg cgccgtcgat ggcgaccgca gttactcagg
     4801 cctcatgacc aaaaaccccg gccacatcgc ctgggaaacg gaatggctcc actcagatct
     4861 ctacacactc agccacatcg aggccgagct cggcgcgaac atgccaccgc cgcgctggcg
     4921 tcagcagacc acgtacaaag cggctccgac gccgctaggg cggaattgcg cactgttcga
     4981 ttccgtcagg ttgtgggcct atcgtcccgc cctcatgcgg atctacctgc cgacccggaa
     5041 cgtggacgga ctcggccgcg cgatctatgc cgagtgccac gcgcgaaacg ccgaatttcc
     5101 gtgcaacgac gtgtgtcccg gaccgctacc ggacagcgag gtccgcgcca tcgccaacag
     5161 catttggcgt tggatcacaa ccaagtcgcg catttgggcg gacgggatcg tggtctacga
     5221 ggccacactc agtgcgcgcc agtcggccat ctcgcggaag ggcgcagcag cgcgcacggc
     5281 ggcgagcaca gttgcgcggc gcgcaaagtc cgcgtcagcc atgcatggag gcattgctat
     5341 gagcgacggc tacagcgacg gctacaaccg gcagccgact gtccgcaaaa agcggcgcgt
     5401 gaccgccgcc gaaggcgctc gaatcaccgg actatccgaa cgccacgtcg tccggctcgt
     5461 ggcgcaggaa cgcagcgagt ggctcgccga gcaggctgca cgccgcgaac gcatccgcgc
     5521 ctatcacgac gacgagggcc actcttggcc gcaaacggcc aaacatttcg ggctgcatct
     5581 ggacaccgtt aagcgactcg gctatcgggc gaggaaagag cgtgcggcag aacaggaagc
     5641 ggctcaaaag gcccacaacg aagccgacaa tccaccgctg ttctaacgca attggggagc
     5701 gggtgtcgcg ggggttccgt ggggggttcc gttgcaacgg gtcggacagg taaaagtcct
     5761 ggtagacgct agttttctgg tttgggccat gcctgtctcg ttgcgtgttt cgttgcgccc
     5821 gttttgaata ccagccagac gagacggggt tctacgaatc ttggtcgata ccaagccatt
     5881 tccgctgaat atcggggagc tcaccgccag aatcggtggt tgtggtgatg tacgtggcga
     5941 actccgttgt agtgcctgtg gtggcatccg tggcgcggcc gcggtaccag atctttaaat
     6001 ctagataaag aagtgacgcg gtctcaagcg tcgagcgtcg ccagcgtgtc gaggatgtcg
     6061 aagtcgtagc cgtcggcgct ggcgatgtag acctgctggt cgaattgact gtcgcgcata
     6121 cacatcgggc cccggggccc gtcgaacccg acatcgtgcg cggatgccat caggtccggt
     6181 atctcggggg agtgggcccg ctggaagatg gcctcgagcg caagcagacc ctcgtaacag
     6241 gattcggcca tcgcgttgag cggtggcgcg tcggcgccgt agcgggcgac gtagctgccc
     6301 atcaggtcca tcgcacccgc ggtggccagt gaactgaagt acgccgcggc gacatagagg
     6361 ttttcggtgg agccggcgcc gctggccagc agcatgttct cctccatcag cgggctgaac
     6421 cgcgccatgc ggtcgtgccc gccggcgcgc gcgaactcgc ggttgaacaa cacggcgtcc
     6481 tggccgacga gcagcatcaa cacggcctgc gcccccgacg cgatggcctt gcggacaggt
     6541 gcgcggaaat cgtcggtgcc gtacgggacg tagatctccc gtctgagctc gaggtccaga
     6601 tctcggcagt acgcgcgggc ggccgcggcg gaacggcgcg gccagatgta gtcatcgccg
     6661 accaggcacc aggaccggat gccgaagtgg tcgcgcagcc aggcgagcgc gggcgcgatc
     6721 tggatctgcg gtgtctcgcc tgtgcagaac acgcccggtg tgcgttcacc gccctcgtac
     6781 aacgaggtgt agacgtacgg gatgcggtcg cggaccaccg gggagatgcg gttgcgcacg
     6841 gccgagatgt gccagccggt cacggcgtcg agaccgtgac ctcgcaaccg gtcggcgacg
     6901 gtccgggcga cgtcgtcgcc gggcgctccg ccgtcgagca cctcgatggt gaccttgcgg
     6961 ccctgcaggc cgcctcggtc gttgacctcc ttggccgcga gctcggccac ggcctcgcac
     7021 gaaggcgcga agattcccgc tggcccttga agcggaatca ccagcccgac gcggaactca
     7081 acctcgccgt cctgcactcc agatcaccgt cgatcccgtg tagtctgcgc ttcaaaggtt
     7141 tctagcagaa ataattcatt ctgaacagac cccgccgtcg acacgaggag acccaacatg
     7201 gccgccggac agcagcgccg ccccaacctc ctgctgccgt tggtgcgtct gacccacctc
     7261 gcggagtcgg cgatcgaacg cgtgctcgcg gactcgtcgc tcaagatcga ggactggcgc
     7321 gtgctcgacg agttggccgg acggcgcacc gtgcccatga gcgatctcgc gcaggccacg
     7381 ctgatcacgg gtccgactct caccagaacc gtcgatcgcc ttgtgtcgca agggatcatc
     7441 taccggactg ccgatctgca tgaccgccgg cgggtgctcg tggcgttgac cccgcggggg
     7501 cggacgctgc gcaaccgcct ggtggacgcg gtagccgagg ccgagtgtgc ggcttttgaa
     7561 tcgtgcgggc tggacgtcga ccagttgcgc gaactcgtcg acaccacctc gaatttgact
     7621 tcgtaaccac ccgcgcccgg cgcgggcgtt cacccttgac ttttattttc atctggatat
     7681 atttcgggtg aatggaaagg ggtgaccatg ccgacctaca cattccgttg ttcccactgc
     7741 ggtcccttcg atctcacctg cgcgatctcc gagcgcgatg cggcggcgac ctgtccggag
     7801 tgccggacgc cggcgcgccg ggtcttcggt tcggtagggc tgacgacatt caccgcggga
     7861 catcaccgcg cattcgacgc ggcgtccgcg agcgccgaaa gtcccacggt ggtgaagtcg
     7921 attcccgcag gcgcggaccg cccgcgggcc ccgcgccgca atcccggtct accgagtctg
     7981 ccgaggtact agcgacatgg gtggcgtcgg gctcatctac gtgggtgcgg tgctcatcat
     8041 cgacgggctg atgctgctgg gccgcatcag cccacgaggc gcaacaccgc tgaacttctt
     8101 cgtcggcgga ctgcaggtgg tgacgcctac ggtgctgatc ctgcagtccg gcggagacgc
     8161 ggccgtgatc ttcgcggcct ccgggctcta cctgttcggc ttcacctacc tgtgggtggc
     8221 catcaacaac gtgaccgact gggacggaga aggtctcgga tggttc
//
