LOCUS       sequence-177496-        5606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLaTM, a genetic tool to measure homo- and heterotypic transmembrane
            helix-helix interactions. Plasmid encoding the 1.1 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 5606)
  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 5606)
  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 1.1
FEATURES             Location/Qualifiers
     source          1..5606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..390
                     /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     313..332
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             603..981
                     /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(637..656)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1961..1978
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2031..2049)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2045..2092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3748..4407)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4322..4341
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4408..4510)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5036..5581)
                     /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(5116..5135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatatttcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
       61 tacgatataa gttgtaattc tcatgtttga cagcttatca tcgataagaa accaattgtc
      121 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac
      181 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa
      241 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg
      301 tcacactttg ctatgccata gcatttttat ccataagatt agcggatgct acctgacgct
      361 ttttatcgca actctctact gtttctccat acccgttttt ttgggctcta gataacgagg
      421 gcaaaaaatg cattataaac tgctgtttgc ggccgcggcc gcgagcctgg cgagcgcggt
      481 gagcgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      541 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      601 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      661 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      721 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      781 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      841 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      901 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgacgcc
      961 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc actagtggca gcggcggcgg
     1021 cagcggcggc ggcagcggtt ctgctaatcg agctagcgtg atcattctgg cggtggggcc
     1081 cggcctgctg ctcctgggga tccacaaact ggcagttaat gttcagctgc ctgccgaagc
     1141 agcagcgaaa gaagccgctg ccaaagaagc ggcagcgaaa gaggctgccg cgaaagaggc
     1201 agcagctaaa gccgcaagca aaggtggaga actgtttacc ggtgttgttc cgattctggt
     1261 tgaactggat ggtgatgtta atggccacaa attctcagtt cgtggtgaag gcgaaggtga
     1321 tgcaaccaat ggtaaactga ccctgaaatt tatctgtacc accggcaaac tgccggttcc
     1381 gtggccgacc ctggttacca ccctgaccta tggtgttcag tgtttcagcc gttatccgga
     1441 tcatatgaaa cagcacgatt tcttcaaatc tgcaatgccg gaaggttatg ttcaagaacg
     1501 taccatctcc tttaaagatg atggcaccta taagacccgt gccgaagtta aatttgaagg
     1561 tgataccctg gtgaatcgca ttgaactgaa aggcatcgat ttcaaagaag atggtaatat
     1621 cctgggccat aaactggaat ataatttcaa cagccacaac gtgtatatca ccgcagataa
     1681 acagaagaat ggcatcaaag ccaactttaa gatccgccat aatgttgaag atggcagcgt
     1741 tcagctggca gatcattatc agcagaatac accgattggt gatggtccgg ttctgctgcc
     1801 ggataatcat tatctgagca cccagagcgt tctgagcaaa gatccgaatg agaaacgtga
     1861 tcacatggtg ctgctggaat ttgttaccgc agcaggtatt acccatggta tggatgaact
     1921 gtataaaggc gattataaag atcacgacgg ctaactcgag caccaccacc accaccactg
     1981 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2041 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaccgatgc
     2101 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2161 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2221 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     2281 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     2341 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     2401 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     2461 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     2521 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc
     2581 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     2641 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     2701 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     2761 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     2821 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     2881 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     2941 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     3001 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     3061 aagtctggaa acgcggaagt cccctacgtg ctgctgaagt tgcccgcaac agagagtgga
     3121 accaaccggt gataccacga tactatgact gagagtcaac gccatgagcg gcctcatttc
     3181 ttattctgag ttacaacagt ccgcaccgct gccggtagct ccttccggtg ggcgcggggc
     3241 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     3301 ccggcagcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     3361 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     3421 acacctacat ctgtattaac gaagcgctaa ccgtttttat caggctctgg gaggcagaat
     3481 aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac tggcctcagg
     3541 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     3601 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt
     3661 cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta
     3721 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     3781 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     3841 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     3901 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     3961 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     4021 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     4081 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     4141 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     4201 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     4261 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     4321 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     4381 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     4441 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     4501 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     4561 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     4621 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     4681 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
     4741 gtaacggcaa aagcaccgcc ggacatcagc gctggcggag tgtatactgg cttactatgt
     4801 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     4861 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     4921 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     4981 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcagcaaag ccgtttttcc
     5041 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     5101 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     5161 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     5221 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     5281 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5341 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     5401 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     5461 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     5521 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     5581 agaagatcat cttattaatc agataa
//
