LOCUS       sequence-415390-        5627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLaTM-System; Antiparallel positive control EmrE_TMD4_3P in NBLa 1.2
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5627)
  AUTHORS   Julius A, Laur L, Schanzenbach C, Langosch D
  TITLE     BLaTM 2.0, a genetic tool revealing preferred antiparallel 
            interaction of transmembrane helix 4 of the dual-topology protein 
            EmrE.
  JOURNAL   J Mol Biol. 2017 Apr 18. pii: S0022-2836(17)30166-3. doi: 
            10.1016/j.jmb.2017.04.003.
  PUBMED    28432015
REFERENCE   2  (bases 1 to 5627)
  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: "J Mol Biol.
            2017 Apr 18. pii: S0022-2836(17)30166-3. doi: 
            10.1016/j.jmb.2017.04.003."
COMMENT     Sequence Label: NBla_1.2_EmrE_TMD4_3P
FEATURES             Location/Qualifiers
     source          1..5627
                     /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             1849..1866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1919..1937)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1933..1980
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3636..4295)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4210..4229
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4296..4398)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4790..4807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4924..5469)
                     /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(5004..5023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5600..5627,1..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 agctagcctg ccggcgatta ttggcatgat gctgatttgc
      961 gcgggcgtgc tgattattaa cctggggatc cacaaactgg cagttaatgt tcagctgcct
     1021 gccgaagcag cagcgaaaga agccgctgcc aaagaagcgg cagcgaaaga ggctgccgcg
     1081 aaagaggcag cagctaaagc cgcaagcaaa ggtggagaac tgtttaccgg tgttgttccg
     1141 attctggttg aactggatgg tgatgttaat ggccacaaat tctcagttcg tggtgaaggc
     1201 gaaggtgatg caaccaatgg taaactgacc ctgaaattta tctgtaccac cggcaaactg
     1261 ccggttccgt ggccgaccct ggttaccacc ctgacctatg gtgttcagtg tttcagccgt
     1321 tatccggatc atatgaaaca gcacgatttc ttcaaatctg caatgccgga aggttatgtt
     1381 caagaacgta ccatctcctt taaagatgat ggcacctata agacccgtgc cgaagttaaa
     1441 tttgaaggtg ataccctggt gaatcgcatt gaactgaaag gcatcgattt caaagaagat
     1501 ggtaatatcc tgggccataa actggaatat aatttcaaca gccacaacgt gtatatcacc
     1561 gcagataaac agaagaatgg catcaaagcc aactttaaga tccgccataa tgttgaagat
     1621 ggcagcgttc agctggcaga tcattatcag cagaatacac cgattggtga tggtccggtt
     1681 ctgctgccgg ataatcatta tctgagcacc cagagcgttc tgagcaaaga tccgaatgag
     1741 aaacgtgatc acatggtgct gctggaattt gttaccgcag caggtattac ccatggtatg
     1801 gatgaactgt ataaaggcga ttataaagat cacgacggct aactcgagca ccaccaccac
     1861 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1921 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1981 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2041 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2101 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     2161 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     2221 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
     2281 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
     2341 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
     2401 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat
     2461 tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc
     2521 atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
     2581 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact
     2641 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg
     2701 cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc
     2761 gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
     2821 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
     2881 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     2941 gtttcgtaaa gtctggaaac gcggaagtcc cctacgtgct gctgaagttg cccgcaacag
     3001 agagtggaac caaccggtga taccacgata ctatgactga gagtcaacgc catgagcggc
     3061 ctcatttctt attctgagtt acaacagtcc gcaccgctgc cggtagctcc ttccggtggg
     3121 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     3181 gacaggtgcc ggcagcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     3241 cgaaacaagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac
     3301 cctgtggaac acctacatct gtattaacga agcgctaacc gtttttatca ggctctggga
     3361 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     3421 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     3481 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     3541 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     3601 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     3661 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     3721 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     3781 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     3841 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     3901 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     3961 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     4021 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     4081 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     4141 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     4201 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     4261 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     4321 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     4381 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     4441 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     4501 tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt
     4561 ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg
     4621 ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tggcggagtg tatactggct
     4681 tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg
     4741 ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg
     4801 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     4861 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cagcaaagcc
     4921 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     4981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     5041 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     5101 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     5161 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5221 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     5281 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     5341 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     5401 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     5461 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
     5521 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
     5581 ttgacagctt atcatcgata agaaaccaat tgtccatatt gcatcag
//
