LOCUS       sequence-417855-        7901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of G protein-coupled receptors for PRESTO-Tango: parallel
            receptorome expression and screening via transcriptional output, 
            with transcriptional activation following arrestin translocation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7901)
  AUTHORS   Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, 
            Sciaky N, Roth BL
  TITLE     PRESTO-Tango as an open-source resource for interrogation of the 
            druggable human GPCRome.
  JOURNAL   Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. 
            Epub 2015 Apr 20.
  PUBMED    25895059
REFERENCE   2  (bases 1 to 7901)
  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/nsmb.3014";
            journalName: "Nat Struct Mol Biol"; date: "2015-05"; volume: "22"; 
            issue: "5"; pages: "362-9"
COMMENT     Sequence Label: HTR1A-Tango
FEATURES             Location/Qualifiers
     source          1..7901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2225..2839,2840..3226)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN
                     KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT
                     EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS
                     MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIEG
                     LLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHADA
                     LDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(2292..2311)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3258..3275)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3264..3488
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3534..3962
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3621..3640)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3831..3852
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3971..3991)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3976..4305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4156..4291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4218..4237
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4372..5166
                     /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(4426..4445)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5036..5055
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5340..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5377..5396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5431..5450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5510..5526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5510..5526)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5523..5545)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5534..5550
                     /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)."
     promoter        complement(join(5558..5564,5565..5582,5583..5588))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5603..5624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5741..5758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5912..6500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5992..6011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6671..7462,7463..7531))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7294..7313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7532..7636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7711..7730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttctgcagat atccagcaca gtggcggccg cgccaccatg aagacgatca tcgccctgag
      781 ctacatcttc tgcctggtat tcgccgacta caaggacgat gatgacgcca gcatcgatat
      841 ggacgtgctg agtcccggcc agggcaacaa caccacttct ccaccggctc cgttcgaaac
      901 tggcggtaat acaaccggca tctctgatgt taccgtcagt tatcaagtga tcactagcct
      961 gctgcttggt actctgatct tctgtgccgt attgggaaac gcatgtgtgg tagcagccat
     1021 tgccctggag cgcagccttc agaatgtggc aaactatttg attggctcac tggcagtgac
     1081 ggacctcatg gtgagcgtgc tcgttcttcc aatggcagct ctctatcagg ttctcaataa
     1141 gtggaccctg ggccaagtga cctgtgacct gtttatcgca ctcgacgtgc tttgttgcac
     1201 ctcttccatt ttgcatctgt gtgccatagc attggatagg tactgggcta tcacagatcc
     1261 gatcgactac gtgaacaaga ggacacctcg gcgggctgcc gctctgatca gcttgacgtg
     1321 gctgattggc tttttgatta gtatacctcc catgcttggg tggaggacac ctgaggatag
     1381 aagcgaccct gacgcgtgta ccatttcaaa ggatcacggc tacaccatct actccacctt
     1441 cggagcattc tatataccac tgttgttgat gttggtgttg tatggacgga tattccgcgc
     1501 cgcgcggttc aggatccgga agactgtaaa gaaagtggaa aagactgggg cggatacccg
     1561 gcatggtgcc tcccccgctc cacagcccaa gaagtccgtg aacggggaga gtgggagcag
     1621 gaactggaga ttgggagtgg aatctaaggc cggaggggca ctgtgtgcca acggtgcagt
     1681 aagacaagga gacgacggtg ctgcacttga agtaatcgag gtgcatagag tggggaatag
     1741 taaagaacac ctgcctctcc cctccgaggc cggaccaact ccatgcgccc cggcctcttt
     1801 tgagcggaaa aacgagagaa atgctgaggc aaagcggaag atggcccttg cccgggaacg
     1861 gaaaacagta aaaaccctgg ggattatcat ggggacattc atcctgtgtt ggctgccctt
     1921 tttcatcgtg gcactcgtgc tgccattttg cgaatcatca tgtcatatgc ccacgcttct
     1981 cggggccatc attaattggc tgggttacag caatagcctg ctgaatcccg tcatctacgc
     2041 ttacttcaac aaggatttcc agaatgcctt taaaaagata attaagtgca agttctgtag
     2101 acaaatcgat accggtggac gcaccccacc cagcctgggt ccccaagatg agtcctgcac
     2161 caccgccagc tcctccctgg ccaaggacac ttcatcgacc ggtgagaacc tgtacttcca
     2221 gctaagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     2281 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     2341 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     2401 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     2461 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     2521 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     2581 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
     2641 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     2701 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     2761 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     2821 taaatgtgaa agtgggtccg cgtacagccg cgcgcgtacg aaaaacaatt acgggtctac
     2881 catcgagggc ctgctcgatc tcccggacga cgacgccccc gaagaggcgg ggctggcggc
     2941 tccgcgcctg tcctttctcc ccgcgggaca cacgcgcaga ctgtcgacgg cccccccgac
     3001 cgatgtcagc ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc
     3061 cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg
     3121 atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga
     3181 gcagatgttt accgatgccc ttggaattga cgagtacggt gggtagtcta gagggcccgt
     3241 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3301 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3361 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3421 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3481 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3541 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3601 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3661 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3721 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3781 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3841 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3901 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3961 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4321 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4381 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4441 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4501 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4561 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4621 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4681 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4741 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4801 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4861 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4921 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4981 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5041 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5101 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5161 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5221 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5281 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5461 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5521 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5581 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5641 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5701 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5821 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7681 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7741 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7801 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     7861 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
