LOCUS       sequence-67685-e        5856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains A2B L980A TMCY mutant, expresses wt AraC for heterodimer 
            competition assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5856)
  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 5856)
  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: pAraTMwt
FEATURES             Location/Qualifiers
     source          1..5856
                     /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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(267..344,345..1442)
                     /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     1416..1439
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1443..1469
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2039..2056)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2209..2295
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2387..2414
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2434..2525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2526..2594,2595..3386)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2744..2763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3557..4145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4046..4065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4299..4316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4331..4471
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4397..4416)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4557..4579
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4657..4734
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4735..5817
                     /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(4754..4773)
                     /label=LacI-R
                     /note="LacI, reverse 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 cagaccatgg aaataaaaac aggtgcacgc atcctcgcat
      301 tatccgcatt aacgacgatg atgttttccg cctcggctct cgccaaaatc gaagaaggta
      361 aactggtaat ctggattaac ggcgataaag gctataacgg tctcgctgaa gtcggtaaga
      421 aattcgagaa agataccgga attaaagtca ccgttgagca tccggataaa ctggaagaga
      481 aattcccaca ggttgcggca actggcgatg gccctgacat tatcttctgg gcacacgacc
      541 gctttggtgg ctacgctcaa tctggcctgt tggctgaaat caccccggac aaagcgttcc
      601 aggacaagct gtatccgttt acctgggatg ccgtacgtta caacggcaag ctgattgctt
      661 acccgatcgc tgttgaagcg ttatcgctga tttataacaa agatctgctg ccgaacccgc
      721 caaaaacctg ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg
      781 cgctgatgtt caacctgcaa gaaccgtact tcacctggcc gctgattgct gctgacgggg
      841 gttatgcgtt caagtatgaa aacggcaagt acgacattaa agacgtgggc gtggataacg
      901 ctggcgcgaa agcgggtctg accttcctgg ttgacctgat taaaaacaaa cacatgaatg
      961 cagacaccga ttactccatc gcagaagctg cctttaataa aggcgaaaca gcgatgacca
     1021 tcaacggccc gtgggcatgg tccaacatcg acaccagcaa agtgaattat ggtgtaacgg
     1081 tactgccgac cttcaagggt caaccatcca aaccgttcgt tggcgtgctg agcgcaggta
     1141 ttaacgccgc cagtccgaac aaagagctgg caaaagagtt cctcgaaaac tatctgctga
     1201 ctgatgaagg tctggaagcg gttaataaag acaaaccgct gggtgccgta gcgctgaagt
     1261 cttacgagga agagttggcg aaagatccac gtattgccgc cactatggaa aacgcccaga
     1321 aaggtgaaat catgccgaac atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg
     1381 cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga
     1441 cttatccgta tgatgtgccg gattatgcgg agctcgcgat tccgatttgg tgggtgctgg
     1501 tgggcgtgct gggcggcctg ctgctggcga ccattctggt gctggcgatg tggaaagtgg
     1561 gcttttttaa acgcaaccgc ccgccgctgg aagaagatga tgaagaaggc gaaggtacca
     1621 tgaacgagtc gctccatcca ccgatggata atcgggtacg cgaggcttgt cagtacatca
     1681 gcgatcacct ggcagacagc aattttgata tcgccagcgt cgcacagcat gtttgcttgt
     1741 cgccgtcgcg tctgtcacat cttttccgcc agcagttagg gattagcgtc ttaagctggc
     1801 gcgaggacca acgcattagt caggcgaagc tgcttttgag cactacccgg atgcctatcg
     1861 ccaccgtcgg tcgcaatgtt ggttttgacg atcaactcta tttctcgcga gtatttaaaa
     1921 aatgcaccgg ggccagcccg agcgagtttc gtgccggttg tgaagaaaaa gtgaatgatg
     1981 tagccgtcaa gttgtcataa aagcttggct gttttggcgg atgagagaag attttcagcc
     2041 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     2101 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     2161 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     2221 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2281 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2341 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     2401 acggatggcc tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa
     2461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2761 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3061 cgatgcctac agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4261 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4321 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4381 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     4441 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4501 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4561 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa
     4621 ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc
     4681 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     4741 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     4801 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     4861 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     4921 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     4981 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     5041 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     5101 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     5161 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     5221 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     5281 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     5341 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     5401 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     5461 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     5521 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     5581 atcccgccgt caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     5641 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     5701 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5761 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5821 caacgcaatt aatgtgagtt agcgcgaatt gatctg
//
