LOCUS       sequence-417784-        5538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLaTM-System GpA wt positive control in CBLa 1.2 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5538)
  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 5538)
  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: CBLa_1.2_GpA_wt
FEATURES             Location/Qualifiers
     source          1..5538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(94..972)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        999..1283
                     /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     1206..1225
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     sig_peptide     1321..1383
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     primer_bind     complement(2694..2711)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2864..2950
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3042..3069
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3237..3256)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3248..4063
                     /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(3321..3340)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4145..4733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4634..4653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4887..4904
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4919..5059
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4985..5004)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5145..5167
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5161..5352)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
       61 cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt
      121 cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa
      181 tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg
      241 catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct
      301 taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca
      361 aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta
      421 ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc
      481 catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc
      541 ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc
      601 ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg
      661 ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg
      721 acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac
      781 aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata
      841 acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg
      901 cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg
      961 cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg
     1021 catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac
     1081 cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa
     1141 caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact
     1201 ttgctatgcc atagcatttt tatccataag attagcggat gctacctgac gctttttatc
     1261 gcaactctct actgtttctc catacccgtt tttttgggct ctagataacg agggcaaaaa
     1321 atgaaaaaga ccgcgattgc gattgcggtg gcgctggcgg gctttgcgac cgtggcgcag
     1381 gcggcgctac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1441 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1501 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1561 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1621 cagatcgctg agataggtgc ctcactgatt aagcattgga ctagtggcag cggcggcggc
     1681 agcggcggcg gcagcggttc tgctaatcga gctagcaccc tgattatttt tggcgtgatg
     1741 gcgggcgtga ttggcaccat tctgctgatt agcgggatcc acaaactggc agttaatgtt
     1801 cagctgcctg ccgaagcagc agcgaaagaa gccgctgcca aagaagcggc agcgaaagag
     1861 gctgccgcga aagaggcagc agctaaagcc gcaagcaaag gtggagaact gtttaccggt
     1921 gttgttccga ttctggttga actggatggt gatgttaatg gccacaaatt ctcagttcgt
     1981 ggtgaaggcg aaggtgatgc aaccaatggt aaactgaccc tgaaatttat ctgtaccacc
     2041 ggcaaactgc cggttccgtg gccgaccctg gttaccaccc tgacctatgg tgttcagtgt
     2101 ttcagccgtt atccggatca tatgaaacag cacgatttct tcaaatctgc aatgccggaa
     2161 ggttatgttc aagaacgtac catctccttt aaagatgatg gcacctataa gacccgtgcc
     2221 gaagttaaat ttgaaggtga taccctggtg aatcgcattg aactgaaagg catcgatttc
     2281 aaagaagatg gtaatatcct gggccataaa ctggaatata atttcaacag ccacaacgtg
     2341 tatatcaccg cagataaaca gaagaatggc atcaaagcca actttaagat ccgccataat
     2401 gttgaagatg gcagcgttca gctggcagat cattatcagc agaatacacc gattggtgat
     2461 ggtccggttc tgctgccgga taatcattat ctgagcaccc agagcgttct gagcaaagat
     2521 ccgaatgaga aacgtgatca catggtgctg ctggaatttg ttaccgcagc aggtattacc
     2581 catggtatgg atgaactgta taaaggcgat tataaagatc acgacggcta actcgaggtc
     2641 gacctgcagg catgcaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     2701 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     2761 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     2821 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     2881 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     2941 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     3001 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     3061 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     3121 atccgctcat ggagctcaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     3181 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     3241 gggtgttatg agccatattc aacgggaaac gtcttgctcc aggccgcgat taaattccaa
     3301 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3361 gacaatctat cgtttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3421 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3481 tatgcctctt ccgtccatca agcattttat ccggactcct gatgaagcat ggttactcac
     3541 cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3601 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3661 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3721 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3781 ctggaaagaa atgcacaaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3841 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3901 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3961 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     4021 gaataaattg cagtttcatt tgatgctcga tgagtttttc taagagctcc atgaccaaaa
     4081 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4141 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4201 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4261 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4321 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4381 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4441 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4501 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4561 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4621 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4681 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4741 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4801 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4861 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4921 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     4981 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     5041 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5101 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5161 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     5221 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     5281 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     5341 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     5401 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     5461 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     5521 tcagggtcaa tgccagcg
//
