LOCUS       sequence-179971-        5609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLaTM, a genetic tool to measure homo- and heterotypic transmembrane
            helix-helix interactions. Plasmid encoding the 1.2 fusionprotein 
            containing the N-terminal fragment of the split beta-lactamase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5609)
  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 5609)
  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: "2017-03-7"; volume: "7:43476. doi"; pages: " 
            10.1038/srep43476"
COMMENT     Sequence Label: N-BLa 1.2
FEATURES             Location/Qualifiers
     source          1..5609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        78..362
                     /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     285..304
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     sig_peptide     400..462
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             578..956
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(612..631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1936..1953
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2006..2024)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2020..2067
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3723..4382)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4297..4316
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4383..4485)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4877..4894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5011..5556)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5091..5110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgttt
       61 gacagcttat catcgataag aaaccaattg tccatattgc atcagacatt gccgtcactg
      121 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
      181 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
      241 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
      301 atccataaga ttagcggatg ctacctgacg ctttttatcg caactctcta ctgtttctcc
      361 atacccgttt ttttgggctc tagataacga gggcaaaaaa tgaaaaagac cgcgattgcg
      421 attgcggtgg cgctggcggg ctttgcgacc gtggcgcagg cggcgccaga aacgctggtg
      481 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      541 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      601 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
      661 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      721 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      781 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      841 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      901 gccataccaa acgacgagcg tgacaccacg acgcctgtag caatggcaac aacgttgcgc
      961 aaactattaa ctggcactag tggcagcggc ggcggcagcg gcggcggcag cggttctgct
     1021 aatcgagcta gcgtgatcat tctggcggtg gggcccggcc tgctgctcct ggggatccac
     1081 aaactggcag ttaatgttca gctgcctgcc gaagcagcag cgaaagaagc cgctgccaaa
     1141 gaagcggcag cgaaagaggc tgccgcgaaa gaggcagcag ctaaagccgc aagcaaaggt
     1201 ggagaactgt ttaccggtgt tgttccgatt ctggttgaac tggatggtga tgttaatggc
     1261 cacaaattct cagttcgtgg tgaaggcgaa ggtgatgcaa ccaatggtaa actgaccctg
     1321 aaatttatct gtaccaccgg caaactgccg gttccgtggc cgaccctggt taccaccctg
     1381 acctatggtg ttcagtgttt cagccgttat ccggatcata tgaaacagca cgatttcttc
     1441 aaatctgcaa tgccggaagg ttatgttcaa gaacgtacca tctcctttaa agatgatggc
     1501 acctataaga cccgtgccga agttaaattt gaaggtgata ccctggtgaa tcgcattgaa
     1561 ctgaaaggca tcgatttcaa agaagatggt aatatcctgg gccataaact ggaatataat
     1621 ttcaacagcc acaacgtgta tatcaccgca gataaacaga agaatggcat caaagccaac
     1681 tttaagatcc gccataatgt tgaagatggc agcgttcagc tggcagatca ttatcagcag
     1741 aatacaccga ttggtgatgg tccggttctg ctgccggata atcattatct gagcacccag
     1801 agcgttctga gcaaagatcc gaatgagaaa cgtgatcaca tggtgctgct ggaatttgtt
     1861 accgcagcag gtattaccca tggtatggat gaactgtata aaggcgatta taaagatcac
     1921 gacggctaac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
     1981 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     2041 tctaaacggg tcttgagggg ttttttgacc gatgcccttg agagccttca acccagtcag
     2101 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2161 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     2221 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     2281 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     2341 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     2401 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     2461 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
     2521 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     2581 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
     2641 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     2701 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
     2761 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc
     2821 agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg
     2881 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     2941 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     3001 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcccct
     3061 acgtgctgct gaagttgccc gcaacagaga gtggaaccaa ccggtgatac cacgatacta
     3121 tgactgagag tcaacgccat gagcggcctc atttcttatt ctgagttaca acagtccgca
     3181 ccgctgccgg tagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     3241 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgcccaac agtcccccgg
     3301 ccacggggcc tgccaccata cccacgccga aacaagcgcc ctgcaccatt atgttccgga
     3361 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc
     3421 gctaaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt caattattac
     3481 ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca cggtcacact
     3541 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     3601 ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta
     3661 ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa
     3721 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     3781 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     3841 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     3901 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     3961 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     4021 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     4081 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     4141 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg
     4201 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     4261 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     4321 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     4381 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     4441 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc
     4501 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg
     4561 aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac
     4621 ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag
     4681 ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca
     4741 tcagcgctgg cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
     4801 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
     4861 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     4921 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     4981 gggaagtgag agggccgcag caaagccgtt tttccatagg ctccgccccc ctgacaagca
     5041 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     5101 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     5161 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     5221 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     5281 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     5341 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     5401 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     5461 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     5521 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     5581 aaatatttct agatttcagt gcaatttat
//
