LOCUS       sequence-211188-        8262 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 8262)
  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 8262)
  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 (Apr)
FEATURES             Location/Qualifiers
     source          1..8262
                     /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"
     CDS             complement(2164..2964)
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      3322..3910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3811..3830
                     /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 aggtaccatg gatcagccgt ccaaatgcgg gattttcgtt agtcggaggc caaacggcat
     2041 tgagcgtcag catatcatca gcgagctgaa gaaagacaat ccccgatccg ctccacgtgt
     2101 tgccccagca atcagcgcga ccttgcccct ccaacgtcat ctcgttctcc gctcatgagc
     2161 tcagccaatc gactggcgag cggcatcgca ttcttcgcat cccgcctctg gcggatgcag
     2221 gaagatcaac ggatctcggc ccagttgacc cagggctgtc gccacaatgt cgcgggagcg
     2281 gatcaaccga gcaaaggcat gaccgactgg accttccttc tgaaggctct tttccttgag
     2341 ccacctgtcc gccaaggcaa agcgctcaca gcagtggtca ttctcgagat aatcgacgcg
     2401 taccaacttg ccatcctgaa gaatggtgca gtgtctcggc accccatagg gaacctttgc
     2461 catcaactcg gcaagatgca gcgtcgtgtt ggcatcgtgt cccacgccga ggagaagtac
     2521 ctgcccatcg agttcatgga cacgggcgac cgggcttgca ggcgagtgag gtggcagggg
     2581 caatggatca gagatgatct gctctgcctg tggccccgct gccgcaaagg caaatggatg
     2641 ggcgctgcgc tttacatttg gcaggcgcca gaatgtgtca gagacaactc caaggtccgg
     2701 tgtaacgggc gacgtggcag gatcgaacgg ctcgtcgtcc agacctgacc acgagggcat
     2761 gacgagcgtc cctcccggac ccagcgcagc acgcagggcc tcgatcagtc caagtggccc
     2821 atcttcgagg ggccggacgc tacggaagga gctgtggacc agcagcacac cgccgggggt
     2881 aaccccaagg ttgagaagct gaccgatgag ctcggctttt cgccattcgt attgcacgac
     2941 attgcactcc accgctgatg acatcagtcg atcatagcac gatcaacggc actgttgcaa
     3001 atagtcggtg gtgataaact tatcatcccc ttttgctgat ggagctgcac atgaaccaaa
     3061 aggatctagg tgaagatcct tctagatacg ccacgtcggg gagtcaggca actatggatg
     3121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3241 tctaggtgaa gatccttttt gataatctca tgctctagtt ccatgagcgt cagaccccgt
     3301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3721 aagcgccacg cttcccgagg ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3901 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3961 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4021 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcaacgcgt gcggccgcac
     4081 gcgtgagccc accagctccg taagttcggg cgctgtgtgg ctcgtacccg cgcattcagg
     4141 cggcaggggg tctaacgggt ctaaggcggc gtgtacggcc gccacagcgg ctctcagcgg
     4201 cccggaaacg tcctcgaaac gacgcatgtg ttcctcctgg ttggtacagg tggttggggg
     4261 tgctcggctg tcgctggtgt tccaccacca gggctcgacg ggagagcggg ggagtgtgca
     4321 gttgtggggt ggcccctcag cgaaatatct gacttggagc tcgtgtcgga ccatacaccg
     4381 gtgattaatc gtggtctact accaagcgtg agccacgtcg ccgacgaatt tgagcagctc
     4441 tggctgccgt actggccgct ggcaagcgac gatctgctcg aggggatcta ccgccaaagc
     4501 cgcgcgtcgg ccctaggccg ccggtacatc gaggcgaacc caacagcgct ggcaaacctg
     4561 ctggtcgtgg acgtagacca tccagacgca gcgctccgag cgctcagcgc ccgggggtcc
     4621 catccgctgc ccaacgcgat cgtgggcaat cgcgccaacg gccacgcaca cgcagtgtgg
     4681 gcactcaacg cccctgttcc acgcaccgaa tacgcgcggc gtaagccgct cgcatacatg
     4741 gcggcgtgcg ccgaaggcct tcggcgcgcc gtcgatggcg accgcagtta ctcaggcctc
     4801 atgaccaaaa accccggcca catcgcctgg gaaacggaat ggctccactc agatctctac
     4861 acactcagcc acatcgaggc cgagctcggc gcgaacatgc caccgccgcg ctggcgtcag
     4921 cagaccacgt acaaagcggc tccgacgccg ctagggcgga attgcgcact gttcgattcc
     4981 gtcaggttgt gggcctatcg tcccgccctc atgcggatct acctgccgac ccggaacgtg
     5041 gacggactcg gccgcgcgat ctatgccgag tgccacgcgc gaaacgccga atttccgtgc
     5101 aacgacgtgt gtcccggacc gctaccggac agcgaggtcc gcgccatcgc caacagcatt
     5161 tggcgttgga tcacaaccaa gtcgcgcatt tgggcggacg ggatcgtggt ctacgaggcc
     5221 acactcagtg cgcgccagtc ggccatctcg cggaagggcg cagcagcgcg cacggcggcg
     5281 agcacagttg cgcggcgcgc aaagtccgcg tcagccatgc atggaggcat tgctatgagc
     5341 gacggctaca gcgacggcta caaccggcag ccgactgtcc gcaaaaagcg gcgcgtgacc
     5401 gccgccgaag gcgctcgaat caccggacta tccgaacgcc acgtcgtccg gctcgtggcg
     5461 caggaacgca gcgagtggct cgccgagcag gctgcacgcc gcgaacgcat ccgcgcctat
     5521 cacgacgacg agggccactc ttggccgcaa acggccaaac atttcgggct gcatctggac
     5581 accgttaagc gactcggcta tcgggcgagg aaagagcgtg cggcagaaca ggaagcggct
     5641 caaaaggccc acaacgaagc cgacaatcca ccgctgttct aacgcaattg gggagcgggt
     5701 gtcgcggggg ttccgtgggg ggttccgttg caacgggtcg gacaggtaaa agtcctggta
     5761 gacgctagtt ttctggtttg ggccatgcct gtctcgttgc gtgtttcgtt gcgcccgttt
     5821 tgaataccag ccagacgaga cggggttcta cgaatcttgg tcgataccaa gccatttccg
     5881 ctgaatatcg gggagctcac cgccagaatc ggtggttgtg gtgatgtacg tggcgaactc
     5941 cgttgtagtg cctgtggtgg catccgtggc gcggccgcgg taccagatct ttaaatctag
     6001 ataaagaagt gacgcggtct caagcgtcga gcgtcgccag cgtgtcgagg atgtcgaagt
     6061 cgtagccgtc ggcgctggcg atgtagacct gctggtcgaa ttgactgtcg cgcatacaca
     6121 tcgggccccg gggcccgtcg aacccgacat cgtgcgcgga tgccatcagg tccggtatct
     6181 cgggggagtg ggcccgctgg aagatggcct cgagcgcaag cagaccctcg taacaggatt
     6241 cggccatcgc gttgagcggt ggcgcgtcgg cgccgtagcg ggcgacgtag ctgcccatca
     6301 ggtccatcgc acccgcggtg gccagtgaac tgaagtacgc cgcggcgaca tagaggtttt
     6361 cggtggagcc ggcgccgctg gccagcagca tgttctcctc catcagcggg ctgaaccgcg
     6421 ccatgcggtc gtgcccgccg gcgcgcgcga actcgcggtt gaacaacacg gcgtcctggc
     6481 cgacgagcag catcaacacg gcctgcgccc ccgacgcgat ggccttgcgg acaggtgcgc
     6541 ggaaatcgtc ggtgccgtac gggacgtaga tctcccgtct gagctcgagg tccagatctc
     6601 ggcagtacgc gcgggcggcc gcggcggaac ggcgcggcca gatgtagtca tcgccgacca
     6661 ggcaccagga ccggatgccg aagtggtcgc gcagccaggc gagcgcgggc gcgatctgga
     6721 tctgcggtgt ctcgcctgtg cagaacacgc ccggtgtgcg ttcaccgccc tcgtacaacg
     6781 aggtgtagac gtacgggatg cggtcgcgga ccaccgggga gatgcggttg cgcacggccg
     6841 agatgtgcca gccggtcacg gcgtcgagac cgtgacctcg caaccggtcg gcgacggtcc
     6901 gggcgacgtc gtcgccgggc gctccgccgt cgagcacctc gatggtgacc ttgcggccct
     6961 gcaggccgcc tcggtcgttg acctccttgg ccgcgagctc ggccacggcc tcgcacgaag
     7021 gcgcgaagat tcccgctggc ccttgaagcg gaatcaccag cccgacgcgg aactcaacct
     7081 cgccgtcctg cactccagat caccgtcgat cccgtgtagt ctgcgcttca aaggtttcta
     7141 gcagaaataa ttcattctga acagaccccg ccgtcgacac gaggagaccc aacatggccg
     7201 ccggacagca gcgccgcccc aacctcctgc tgccgttggt gcgtctgacc cacctcgcgg
     7261 agtcggcgat cgaacgcgtg ctcgcggact cgtcgctcaa gatcgaggac tggcgcgtgc
     7321 tcgacgagtt ggccggacgg cgcaccgtgc ccatgagcga tctcgcgcag gccacgctga
     7381 tcacgggtcc gactctcacc agaaccgtcg atcgccttgt gtcgcaaggg atcatctacc
     7441 ggactgccga tctgcatgac cgccggcggg tgctcgtggc gttgaccccg cgggggcgga
     7501 cgctgcgcaa ccgcctggtg gacgcggtag ccgaggccga gtgtgcggct tttgaatcgt
     7561 gcgggctgga cgtcgaccag ttgcgcgaac tcgtcgacac cacctcgaat ttgacttcgt
     7621 aaccacccgc gcccggcgcg ggcgttcacc cttgactttt attttcatct ggatatattt
     7681 cgggtgaatg gaaaggggtg accatgccga cctacacatt ccgttgttcc cactgcggtc
     7741 ccttcgatct cacctgcgcg atctccgagc gcgatgcggc ggcgacctgt ccggagtgcc
     7801 ggacgccggc gcgccgggtc ttcggttcgg tagggctgac gacattcacc gcgggacatc
     7861 accgcgcatt cgacgcggcg tccgcgagcg ccgaaagtcc cacggtggtg aagtcgattc
     7921 ccgcaggcgc ggaccgcccg cgggccccgc gccgcaatcc cggtctaccg agtctgccga
     7981 ggtactagcg acatgggtgg cgtcgggctc atctacgtgg gtgcggtgct catcatcgac
     8041 gggctgatgc tgctgggccg catcagccca cgaggcgcaa caccgctgaa cttcttcgtc
     8101 ggcggactgc aggtggtgac gcctacggtg ctgatcctgc agtccggcgg agacgcggcc
     8161 gtgatcttcg cggcctccgg gctctacctg ttcggcttca cctacctgtg ggtggccatc
     8221 aacaacgtga ccgactggga cggagaaggt ctcggatggt tc
//
