LOCUS       sequence-172660-        5610 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 5610)
  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 5610)
  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..5610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..199
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     sig_peptide     295..357
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             473..851
                     /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(507..526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1831..1848
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1901..1919)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1915..1962
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3618..4277)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4192..4211
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4278..4380)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4772..4789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4906..5451)
                     /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(4986..5005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5583..5610,1..257)
                     /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)"
ORIGIN
        1 acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct
       61 tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag
      121 tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta
      181 tgccatagca tttttatcca taagattagc ggatgctacc tgacgctttt tatcgcaact
      241 ctctactgtt tctccatacc cgtttttttg ggctctagat aacgagggca aaaaatgaaa
      301 aagaccgcga ttgcgattgc ggtggcgctg gcgggctttg cgaccgtggc gcaggcggcg
      361 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      421 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      481 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
      541 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      601 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      661 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
      721 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
      781 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgacgcc tgtagcaatg
      841 gcaacaacgt tgcgcaaact attaactggc actagtggca gcggcggcgg cagcggcggc
      901 ggcagcggtt ctgctaatcg agctagcgtg atcattctgg cggtggggcc cggcctgctg
      961 ctcctgggga tccacaaact ggcagttaat gttcagctgc ctgccgaagc agcagcgaaa
     1021 gaagccgctg ccaaagaagc ggcagcgaaa gaggctgccg cgaaagaggc agcagctaaa
     1081 gccgcaagca aaggtggaga actgtttacc ggtgttgttc cgattctggt tgaactggat
     1141 ggtgatgtta atggccacaa attctcagtt cgtggtgaag gcgaaggtga tgcaaccaat
     1201 ggtaaactga ccctgaaatt tatctgtacc accggcaaac tgccggttcc gtggccgacc
     1261 ctggttacca ccctgaccta tggtgttcag tgtttcagcc gttatccgga tcatatgaaa
     1321 cagcacgatt tcttcaaatc tgcaatgccg gaaggttatg ttcaagaacg taccatctcc
     1381 tttaaagatg atggcaccta taagacccgt gccgaagtta aatttgaagg tgataccctg
     1441 gtgaatcgca ttgaactgaa aggcatcgat ttcaaagaag atggtaatat cctgggccat
     1501 aaactggaat ataatttcaa cagccacaac gtgtatatca ccgcagataa acagaagaat
     1561 ggcatcaaag ccaactttaa gatccgccat aatgttgaag atggcagcgt tcagctggca
     1621 gatcattatc agcagaatac accgattggt gatggtccgg ttctgctgcc ggataatcat
     1681 tatctgagca cccagagcgt tctgagcaaa gatccgaatg agaaacgtga tcacatggtg
     1741 ctgctggaat ttgttaccgc agcaggtatt acccatggta tggatgaact gtataaaggc
     1801 gattataaag atcacgacgg ctaactcgag caccaccacc accaccactg agatccggct
     1861 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     1921 taaccccttg gggcctctaa acgggtcttg aggggttttt tgaccgatgc ccttgagagc
     1981 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     2041 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     2101 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     2161 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     2221 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt
     2281 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     2341 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     2401 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc tgatcgtcac
     2461 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     2521 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     2581 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat
     2641 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg
     2701 tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg
     2761 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     2821 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     2881 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     2941 acgcggaagt cccctacgtg ctgctgaagt tgcccgcaac agagagtgga accaaccggt
     3001 gataccacga tactatgact gagagtcaac gccatgagcg gcctcatttc ttattctgag
     3061 ttacaacagt ccgcaccgct gtccggtagc tccttccggt gggcgcgggg catgactatc
     3121 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     3181 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgccctgc
     3241 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3301 tctgtattaa cgaagcgcta accgttttta tcaggctctg ggaggcagaa taaatgatca
     3361 tatcgtcaat tattacctcc acggggagag cctgagcaaa ctggcctcag gcatttgaga
     3421 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
     3481 cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt tcgaatttct
     3541 gccattcatc cgcttattat cacttattca ggcgtagcac caggcgttta agggcaccaa
     3601 taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     3661 ttaagcattc tgccgacatg gaagccatca cagacggcat gatgaacctg aatcgccagc
     3721 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     3781 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     3841 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     3901 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     3961 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     4021 tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg agcattcatc
     4081 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     4141 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     4201 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     4261 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     4321 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     4381 acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga
     4441 tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg
     4501 ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc
     4561 ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa
     4621 aagcaccgcc ggacatcagc gctggcggag tgtatactgg cttactatgt tggcactgat
     4681 gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc
     4741 agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
     4801 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
     4861 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
     4921 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     4981 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
     5041 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
     5101 cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
     5161 agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg
     5221 caaaagcacc actggcagca gccactggta attgatttag aggagttagt cttgaagtca
     5281 tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag
     5341 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     5401 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     5461 cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca
     5521 cctgaagtca gccccatacg atataagttg taattctcat gtttgacagc ttatcaatcg
     5581 ataagaaacc aattgtccat attgcatcag
//
