LOCUS       sequence-67699-e        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains A2B L980A TMCY mutant, expresses DN AraC (AraC*) for 
            heterodimer competition assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  AUTHORS   Su PC, Berger BW
  TITLE     A Novel Assay for Assessing Juxtamembrane and Transmembrane Domain 
            Interactions Important for Receptor Heterodimerization.
  JOURNAL   J Mol Biol. 2013 Jul 19. pii: S0022-2836(13)00456-7. doi: 
            10.1016/j.jmb.2013.07.022.
  PUBMED    23876708
REFERENCE   2  (bases 1 to 6001)
  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.
            2013 Jul 19. pii: S0022-2836(13)00456-7. doi: 
            10.1016/j.jmb.2013.07.022."
COMMENT     Sequence Label: pAraTMDN
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             385..1467
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(404..423)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1699..1704,1705..1721,1722..1728)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1736..1752
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             join(1773..1850,1851..2948)
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="MEIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2922..2945
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2949..2978
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3542..3559)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3712..3798
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3890..3917
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(4276..5091)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4276..5091)
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4999..5018
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5092..5183)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5199..5948)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(5209..5226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5449..5468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaaattcg agctcggtac ccggggatcc tctagagtcg
      301 acctgcaggc atgccgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
      361 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      421 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      481 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      541 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      601 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      661 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      721 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      781 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      841 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      901 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      961 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     1021 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     1081 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     1141 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     1201 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
     1261 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1321 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1381 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1441 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt
     1501 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1561 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1621 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1681 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
     1741 agcggataac aatttcacac aggaaacaga ccatggaaat aaaaacaggt gcacgcatcc
     1801 tcgcattatc cgcattaacg acgatgatgt tttccgcctc ggctctcgcc aaaatcgaag
     1861 aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc gctgaagtcg
     1921 gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg gataaactgg
     1981 aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc ttctgggcac
     2041 acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc ccggacaaag
     2101 cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac ggcaagctga
     2161 ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat ctgctgccga
     2221 acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa gcgaaaggta
     2281 agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg attgctgctg
     2341 acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg
     2401 ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa aacaaacaca
     2461 tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc gaaacagcga
     2521 tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg aattatggtg
     2581 taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc gtgctgagcg
     2641 caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc gaaaactatc
     2701 tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt gccgtagcgc
     2761 tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccact atggaaaacg
     2821 cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc
     2881 gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg
     2941 cgcagactga acagaaactg attagcgaag aagatctgga gctcgcgatt ccgatttggt
     3001 gggtgctggt gggcgtgctg ggcggcctgc tgctggcgac cattctggtg ctggcgatgt
     3061 ggaaagtggg cttttttaaa cgcaaccgcc cgccgctgga agaagatgat gaagaaggcg
     3121 aaggtaccat gaacgagtcg ctccatccac cgatggataa tcgggtacgc gaggcttgtc
     3181 agtacatcag cgatcacctg gcagacagca attttgatat cgccagcgtc gcacagcatg
     3241 tttgcttgtc gccgtcggct ctgtcacatc ttttccgcca gcagttaggg attagcgtct
     3301 taagctggcg cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc accacccgga
     3361 tgcctatcgc caccgtcggt cgcaatgttg gttttgacga tcaactctat ttctcgcggg
     3421 tatttaaaaa atgcaccggg gccagcccga gcgagttccg tgccggttgt gaagaaaaag
     3481 tgaatgatgt agccgtcaag ttgtcataag gctgttttgg cggatgagag aagattttca
     3541 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     3601 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     3661 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     3721 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     3781 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     3841 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     3901 ctgacggatg gcctttttgc gtttctacaa actctttttg tttatttttc taaatacatt
     3961 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4021 gactagtggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
     4081 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     4141 gaccctgccc tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg cacttttcgg
     4201 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4261 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     4321 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4381 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4441 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4501 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     4561 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     4621 attcattcgt gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt
     4681 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4741 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     4801 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4861 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     4921 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     4981 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     5041 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct
     5101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5161 atgtatttag aaaaataaac aaataggcat gcagcgctct tccgcttcct cgctcactga
     5221 ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca gctcactcaa aagcggtaat
     5281 acggttatcc acagaatcag gggataaagc cggaaagaac atgtgagcaa aaagcaaagc
     5341 accggaagaa gccaacgccg caggcgtttt tccataggct ccgcccccct gacgagcatc
     5401 acaaaaatcg acgctcaagc cagaggtggc gaaacccgac aggactataa agataccagg
     5461 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5521 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgttggt
     5581 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5641 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5701 acttatcgcc actggcagca gccattggta actgatttag aggactttgt cttgaagtta
     5761 tgcacctgtt aaggctaaac tgaaagaaca gattttggtg agtgcggtcc tccaacccac
     5821 ttaccttggt tcaaagagtt ggtagctcag cgaaccttga gaaaaccacc gttggtagcg
     5881 gtggtttttc tttatttatg agatgatgaa tcaatcggtc tatcaagtca acgaacagct
     5941 attccgttac tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcact
     6001 a
//
