LOCUS       sequence-177504-        5517 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 C-terminal fragment of the split beta-lactamase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5517)
  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 5517)
  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: C-BLa 1.1
FEATURES             Location/Qualifiers
     source          1..5517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(79..957)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        984..1268
                     /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     1191..1210
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2658..2675)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2828..2914
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3006..3033
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3201..3220)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3212..4027
                     /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(3285..3304)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4109..4697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4598..4617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4851..4868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4883..5023
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4949..4968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5109..5131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5125..5316)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat
       61 tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca
      121 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
      181 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
      241 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
      301 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
      361 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
      421 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
      481 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
      541 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
      601 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
      661 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
      721 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
      781 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
      841 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
      901 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
      961 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
     1021 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
     1081 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
     1141 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
     1201 atttttatcc ataagattag cggatgctac ctgacgcttt ttatcgcaac tctctactgt
     1261 ttctccatac ccgttttttt gggctctaga taacgagggc aaaaaatgca ttataaactg
     1321 ctgtttgcgg ccgcggccgc gagcctggcg agcgcggtga gcgctcacct acttactcta
     1381 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1441 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1501 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1561 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1621 gcctcactga ttaagcattg gactagtggc agcggcggcg gcagcggcgg cggcagcggt
     1681 tctgctaatc gagctagcgt gatcattctg gcggtggggc ccggcctgct gctcctgggg
     1741 atccacaaac tggcagttaa tgttcagctg cctgccgaag cagcagcgaa agaagccgct
     1801 gccaaagaag cggcagcgaa agaggctgcc gcgaaagagg cagcagctaa agccgcaagc
     1861 aaaggtggag aactgtttac cggtgttgtt ccgattctgg ttgaactgga tggtgatgtt
     1921 aatggccaca aattctcagt tcgtggtgaa ggcgaaggtg atgcaaccaa tggtaaactg
     1981 accctgaaat ttatctgtac caccggcaaa ctgccggttc cgtggccgac cctggttacc
     2041 accctgacct atggtgttca gtgtttcagc cgttatccgg atcatatgaa acagcacgat
     2101 ttcttcaaat ctgcaatgcc ggaaggttat gttcaagaac gtaccatctc ctttaaagat
     2161 gatggcacct ataagacccg tgccgaagtt aaatttgaag gtgataccct ggtgaatcgc
     2221 attgaactga aaggcatcga tttcaaagaa gatggtaata tcctgggcca taaactggaa
     2281 tataatttca acagccacaa cgtgtatatc accgcagata aacagaagaa tggcatcaaa
     2341 gccaacttta agatccgcca taatgttgaa gatggcagcg ttcagctggc agatcattat
     2401 cagcagaata caccgattgg tgatggtccg gttctgctgc cggataatca ttatctgagc
     2461 acccagagcg ttctgagcaa agatccgaat gagaaacgtg atcacatggt gctgctggaa
     2521 tttgttaccg cagcaggtat tacccatggt atggatgaac tgtataaagg cgattataaa
     2581 gatcacgacg gctaactcga ggtcgacctg caggcatgca agcttggctg ttttggcgga
     2641 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     2701 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     2761 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     2821 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2881 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     2941 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     3001 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     3061 ttttctaaat acattcaaat atgtatccgc tcatggagct caaagccacg ttgtgtctca
     3121 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     3181 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     3241 ctccaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3301 cgataatgtc gggcaatcag gtgcgacaat ctatcgtttg tatgggaagc ccgatgcgcc
     3361 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3421 cagactaaac tggctgacgg aatttatgcc tcttccgtcc atcaagcatt ttatccggac
     3481 tcctgatgaa gcatggttac tcaccactgc gatccccggc aaaacagcat tccaggtatt
     3541 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3601 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3661 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3721 taatggctgg cctgttgaac aagtctggaa agaaatgcac aaacttttgc cattctcacc
     3781 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3841 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3901 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3961 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     4021 tttctaagag ctccatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4801 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4861 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4921 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4981 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     5041 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5101 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5161 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5221 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5281 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     5341 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     5401 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     5461 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcgatgcat aatgtgc
//
