LOCUS       sequence-181586-        5251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The C-BLa 2.0 plasmid encodes a protein, where the TMD is connected 
            to the C-terminus of the cytoplasmic domain of the ToxR protein and 
            to the N-terminus of the C-terminal beta-lactamase fragment..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  AUTHORS   Julius A, Laur L, Schanzenbach C, Langosch D
  TITLE     BLaTM 2.0, a genetic tool revealing preferred antiparallel 
            interaction of transmembrane helix 4 of the dual-topology protein 
            EmrE.
  JOURNAL   J Mol Biol. 2017 Apr 18. pii: S0022-2836(17)30166-3. doi: 
            10.1016/j.jmb.2017.04.003.
  PUBMED    28432015
REFERENCE   2  (bases 1 to 5251)
  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: "J Mol Biol.
            2017 Apr 18. pii: S0022-2836(17)30166-3. doi: 
            10.1016/j.jmb.2017.04.003."
COMMENT     Sequence Label: C-BLa 2.0
FEATURES             Location/Qualifiers
     source          1..5251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(44..922)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        949..1233
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1156..1175
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2241..2270
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2274..2291
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2357..2374)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2527..2613
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2705..2732
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2900..2919)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2911..3726
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPSIKHFIRTPDEAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2984..3003)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3808..4396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4297..4316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4550..4567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4582..4722
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4648..4667)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4808..4830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4824..5015)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg
       61 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc
      121 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg
      181 tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct
      241 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg
      301 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat
      361 cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt
      421 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg
      481 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg
      541 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa
      601 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag
      661 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg
      721 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat
      781 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg
      841 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg
      901 gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt
      961 gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca
     1021 aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa
     1081 aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg
     1141 gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat gctacctgac
     1201 gctttttatc gcaactctct actgtttctc catacccgtt tttttgggct ctagataacg
     1261 agggcaaaaa atgttcggat taggacacaa ctcaaaagag atttcgatga gtcatattgg
     1321 tactaaattc attcttgctg aaaaatttac cttcgatccc ctaagcaata ctctgattga
     1381 caaagaagat agtgaagaga tcattcgatt aggcagcaac gaaagccgaa ttctttggct
     1441 gctggcccaa cgtccaaacg aggtgatttc tcgcaatgat ttgcatgact ttgtttggcg
     1501 agagcaaggt tttgaagtcg atgattccag cttaacccaa gccatttcga ctctgcgcaa
     1561 aatgctcaaa gattcgacaa agtccccaca atacgtcaaa acggttccga agcgcggtta
     1621 ccaattgatc gcccgagtgg aaacggttga agaagagatg gctcgcgaaa acgaagctgc
     1681 tcatgacatc tctcagccag aatctgtcaa tgaatacgca gaatcaagca gtgtgccttc
     1741 atcagccact gtagtgaaca caccgcagcc agccaatgtc gtggcgaata aatcggctcc
     1801 aaacttgggg aatcgagcta gcgtgatcat tctggcggtg gggcccggcc tgctgctcct
     1861 ggggatcctg atcaacccaa gccaatccag ctttaaaccc ctaacggatg gtggtagcgg
     1921 cggcagccta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1981 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2041 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2101 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2161 acagatcgct gagataggtg cctcactgat taagcattgg ggcgattata aagatcacga
     2221 cggctaagag ctcggtaccc gaacagaagc tgatcagcga agaagatctg aaccaccacc
     2281 accaccacca ctaactcgag gtcgacctgc aggcatgcaa gcttggctgt tttggcggat
     2341 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     2401 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     2461 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     2521 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2581 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     2641 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     2701 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     2761 tttctaaata cattcaaata tgtatccgct catggagctc aaagccacgt tgtgtctcaa
     2821 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     2881 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     2941 tccaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     3001 gataatgtcg ggcaatcagg tgcgacaatc tatcgtttgt atgggaagcc cgatgcgcca
     3061 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     3121 agactaaact ggctgacgga atttatgcct cttccgtcca tcaagcattt tatccggact
     3181 cctgatgaag catggttact caccactgcg atccccggca aaacagcatt ccaggtatta
     3241 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     3301 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     3361 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     3421 aatggctggc ctgttgaaca agtctggaaa gaaatgcaca aacttttgcc attctcaccg
     3481 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3541 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3601 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3661 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3721 ttctaagagc tccatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4441 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4501 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4561 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4621 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4681 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4741 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4801 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4861 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4921 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4981 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     5041 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5101 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5161 cggcgggacc agagaaaaat cactcagggt caatgccagc gcgatgcata atgtgcctgt
     5221 caaatggacg aagcagggat tctgcaaacc c
//
