LOCUS       sequence-177513-        5520 bp    DNA     circular SYN 14-APR-2025
DEFINITION  BLaTM, a genetic tool to measure homo- and heterotypic transmembrane
            helix-helix interactions. Plasmid encoding the 1.2 fusionprotein 
            containing the C-terminal fragment of the split beta-lactamase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  AUTHORS   Schanzenbach C, Schmidt FC, Breckner P, Teese MG, Langosch D
  TITLE     Identifying ionic interactions within a membrane using BLaTM, a 
            genetic tool to measure homo- and heterotypic transmembrane 
            helix-helix interactions.
  JOURNAL   Sci Rep. 2017 Mar 7;7:43476. doi: 10.1038/srep43476.
  PUBMED    28266525
REFERENCE   2  (bases 1 to 5520)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2017-03-7"; volume: "7:43476. doi"; pages: " 
            10.1038/srep43476"
COMMENT     Sequence Label: C-BLa 1.2
FEATURES             Location/Qualifiers
     source          1..5520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(204..1082)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1109..1393
                     /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     1316..1335
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     sig_peptide     1431..1493
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     primer_bind     complement(2786..2803)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2956..3042
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3134..3161
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3329..3348)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3340..4155
                     /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(3413..3432)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4237..4825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4726..4745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4979..4996
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5011..5151
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5077..5096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5237..5259
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5253..5444)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
       61 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cgatgcataa
      121 tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg
      181 tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct
      241 tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag
      301 aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg
      361 gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta
      421 atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt
      481 gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc
      541 tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc
      601 agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc
      661 ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg
      721 cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg
      781 cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag
      841 tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt
      901 ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt
      961 cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc
     1021 gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc
     1081 atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat
     1141 tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt
     1201 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc
     1261 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc
     1321 atagcatttt tatccataag attagcggat gctacctgac gctttttatc gcaactctct
     1381 actgtttctc catacccgtt tttttgggct ctagataacg agggcaaaaa atgaaaaaga
     1441 ccgcgattgc gattgcggtg gcgctggcgg gctttgcgac cgtggcgcag gcggcgctac
     1501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1741 agataggtgc ctcactgatt aagcattgga ctagtggcag cggcggcggc agcggcggcg
     1801 gcagcggttc tgctaatcga gctagcgtga tcattctggc ggtggggccc ggcctgctgc
     1861 tcctggggat ccacaaactg gcagttaatg ttcagctgcc tgccgaagca gcagcgaaag
     1921 aagccgctgc caaagaagcg gcagcgaaag aggctgccgc gaaagaggca gcagctaaag
     1981 ccgcaagcaa aggtggagaa ctgtttaccg gtgttgttcc gattctggtt gaactggatg
     2041 gtgatgttaa tggccacaaa ttctcagttc gtggtgaagg cgaaggtgat gcaaccaatg
     2101 gtaaactgac cctgaaattt atctgtacca ccggcaaact gccggttccg tggccgaccc
     2161 tggttaccac cctgacctat ggtgttcagt gtttcagccg ttatccggat catatgaaac
     2221 agcacgattt cttcaaatct gcaatgccgg aaggttatgt tcaagaacgt accatctcct
     2281 ttaaagatga tggcacctat aagacccgtg ccgaagttaa atttgaaggt gataccctgg
     2341 tgaatcgcat tgaactgaaa ggcatcgatt tcaaagaaga tggtaatatc ctgggccata
     2401 aactggaata taatttcaac agccacaacg tgtatatcac cgcagataaa cagaagaatg
     2461 gcatcaaagc caactttaag atccgccata atgttgaaga tggcagcgtt cagctggcag
     2521 atcattatca gcagaataca ccgattggtg atggtccggt tctgctgccg gataatcatt
     2581 atctgagcac ccagagcgtt ctgagcaaag atccgaatga gaaacgtgat cacatggtgc
     2641 tgctggaatt tgttaccgca gcaggtatta cccatggtat ggatgaactg tataaaggcg
     2701 attataaaga tcacgacggc taactcgagg tcgacctgca ggcatgcaag cttggctgtt
     2761 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     2821 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     2881 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     2941 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     3001 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     3061 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     3121 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     3181 ttgtttattt ttctaaatac attcaaatat gtatccgctc atggagctca aagccacgtt
     3241 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     3301 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     3361 acgtcttgct ccaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     3421 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgtttgta tgggaagccc
     3481 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     3541 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgtccat caagcatttt
     3601 atccggactc ctgatgaagc atggttactc accactgcga tccccggcaa aacagcattc
     3661 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     3721 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     3781 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     3841 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcacaa acttttgcca
     3901 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3961 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     4021 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     4081 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     4141 gatgagtttt tctaagagct ccatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4381 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4921 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4981 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     5041 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     5101 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     5161 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5221 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5281 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     5341 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     5401 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     5461 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
//
