LOCUS       sequence-353385-        7942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tango assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7942)
  AUTHORS   Yan Y, Xu TH, Melcher K, Xu HE
  TITLE     Defining the minimum substrate and charge recognition model of 
            gamma-secretase.
  JOURNAL   Acta Pharmacol Sin. 2017 Oct;38(10):1412-1424. doi: 
            10.1038/aps.2017.35. Epub 2017 Apr 17.
  PUBMED    28414207
REFERENCE   2  (bases 1 to 7942)
  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"; doi: 
            "10.1038/aps.2017.35"; journalName: "Acta Pharmacol Sin"; date: 
            "2017-10"; volume: "38"; issue: "10"; pages: "1412-1424"
COMMENT     Sequence Label: pCDNA3-IgL-C99-T4L-rTA
FEATURES             Location/Qualifiers
     source          1..7942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(365..387)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     528..547
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        650..1029
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1030..1233
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1184..1204
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1230..1254
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(1339..1383,1466..1477)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGVHS"
     CDS             join(1339..1384,1467..1477)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGVHS"
     primer_bind     1449..1468
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             join(2261..2878,2879..3265)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the reverse tetracycline repressor 
                     rTetR with the C-terminal activation domain of herpes 
                     simplex virus VP16"
                     /label=rtTA
                     /note="In the Tet-On(R) system, rtTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHSDTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(2331..2350)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3465..3484)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3578..3595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3749..4337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3829..3848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4508..5299,5300..5368))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5131..5150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5369..5473)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5541..5559
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5586..5720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5610..5629)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5664..5683
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6145..6165)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          6295..6360
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(6780..7574)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6891..6910)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7501..7520
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7935..7942,1..322))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gctgcctcgc gcgtttcggt gatgacggtg
      361 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      421 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
      481 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
      541 gattgtactg agagtgcacc tctcgaggag cttggcccat tgcatacgtt gtatccatat
      601 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
      661 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      721 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      781 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      841 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      901 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      961 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     1021 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     1081 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     1141 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     1201 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     1261 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
     1321 ggccccgtcg acgccaccat gggatggagc tgtatcatcc tcttcttggt agcaacagct
     1381 acaggtaagg ggctcacagt agcaggcttg aggtctggac atatatatgg gtgacaatga
     1441 catccacttt gcctttctct ccacaggtgt ccactccggt accgacgccg aatttcgcca
     1501 cgacagcggt tacgaagtcc atcatcaaaa actggtgttc tttgcggaag acgttggtag
     1561 caataaaggc gcaattatcg gcctgatggt cggcggtgtg gttattgcga ccgttattgt
     1621 catcacgctg gttatgctga agaaaaaaca gtataccagc atccatcacg gtgtcgtgga
     1681 agtggatgcg gccgttacgc cggaagaacg tcatctgtct aaaatgcaac aaaatggcta
     1741 tgaaaatccg acctacaaat tcttcgaaca gatgcaaaac aacatcttcg aaatgctccg
     1801 gatcgatgag ggactgagac tgaaaatcta caaggacacc gagggctact acacaatcgg
     1861 aatcggacat ctgctcacca aatcaccttc cctgaatgct gccaaatccg agctggacaa
     1921 ggcaattgga cggaatacaa atggagtgat caccaaggat gaggccgaga aactgttcaa
     1981 tcaggacgtg gacgctgctg tgagaggcat tctgagaaat gccaaactca aacccgtgta
     2041 cgactctctc gacgctgtcc ggcgagctgc tctcatcaat atggtgttcc agatgggcga
     2101 aaccggagtc gctggcttta ccaactccct ccggatgctc cagcagaaac gatgggacga
     2161 ggctgctgtg aatctggcca aatctcggtg gtacaaccag acccccaatc gggctaaacg
     2221 agtgattaca accttccgga ccggaacatg ggatgcctac tctagattag ataaaagtaa
     2281 agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg
     2341 taaactcgcc cagaagcttg gtgtagagca gcctacactg tattggcatg taaaaaataa
     2401 gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg
     2461 ccctttagaa ggggaaagct ggcaagattt tttacgcaat aacgctaaaa gttttagatg
     2521 tgctttacta agtcatcgcg atggagcaaa agtacattca gatacacggc ctacagaaaa
     2581 acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga
     2641 gaacgcgtta tatgcactca gcgctgtggg gcattttact ttaggttgcg tattggaaga
     2701 tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc
     2761 attattacga caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt
     2821 cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtccgc
     2881 gtacagccgc gcgcgtacga aaaacaatta cgggtctacc atcgagggcc tgctcgatct
     2941 cccggacgac gacgcccccg aagaggcggg gctggcggct ccgcgcctgt cctttctccc
     3001 cgcgggacac acgcgcagac tgtcgacggc ccccccgacc gatgtcagcc tgggggacga
     3061 gctccactta gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga
     3121 tctggacatg ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc
     3181 cccctacggc gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct
     3241 tggaattgac gagtacggtg ggtagggggc gcgaggatca acatcgataa aataaaagat
     3301 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     3361 agagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     3421 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     3481 ttgctcaatg tacctataac cagaccgttc agctgcatta atgaatcggc caacgcgcgg
     3541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5101 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     5161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5341 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5461 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5521 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cactcgaggt
     5581 cgagggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5641 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5701 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     5761 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     5821 atgatctcta gtcaaggcac tatacatcaa atattcctta ttaacccctt tacaaattaa
     5881 aaagctaaag gtacacaatt tttgagcata gttattaata gcagacactc tatgcctgtg
     5941 tggagtaaga aaaaacagta tgttatgatt ataactgtta tgcctactta taaaggttac
     6001 agaatatttt tccataattt tcttgtatag cagtgcagct ttttcctttg tggtgtaaat
     6061 agcaaagcaa gcaagagttc tattactaaa cacagcatga ctcaaaaaac ttagcaattc
     6121 tgaaggaaag tccttggggt cttctacctt tctcttcttt tttggaggag tagaatgttg
     6181 agagtcagca gtagcctcat catcactaga tggcatttct tctgagcaaa acaggttttc
     6241 ctcattaaag gcattccacc actgctccca ttcatcagtt ccataggttg gaatctaaaa
     6301 tacacaaaca attagaatca gtagtttaac acattataca cttaaaaatt ttatatttac
     6361 cttagagctt taaatctctg taggtagttt gtccaattat gtcacaccac agaagtaagg
     6421 ttccttcaca aagatccgga ccaaagcggc catcgtgcct ccccactcct gcagttcggg
     6481 ggcatggatg cgcggatagc cgctgctggt ttcctggatg ccgacggatt tgcactgccg
     6541 gtagaactcc gcgaggtcgt ccagcctcag gcagcagctg aaccaactcg cgaggggatc
     6601 gagcccgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     6661 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     6721 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     6781 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     6841 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     6901 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     6961 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     7021 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     7081 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     7141 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     7201 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     7261 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     7321 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     7381 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     7441 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     7501 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     7561 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct tgatcccctg
     7621 cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc
     7681 ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag
     7741 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     7801 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     7861 ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag
     7921 cgtgaagcta gctttttgca aa
//
