LOCUS       sequence-417844-        7652 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 7652)
  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 7652)
  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: P2RY10-Tango
FEATURES             Location/Qualifiers
     source          1..7652
                     /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(1976..2590,2591..2977)
                     /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(2043..2062)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3009..3026)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3015..3239
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3285..3713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3372..3391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3582..3603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3722..3742)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3727..4056
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3907..4042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3969..3988
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4123..4917
                     /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(4177..4196)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4787..4806
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5091..5212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5128..5147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5182..5201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5261..5277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5261..5277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5274..5296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5285..5301
                     /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(5309..5315,5316..5333,5334..5339))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5354..5375
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5492..5509)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5663..6251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5743..5762)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6422..7213,7214..7282))
                     /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     7045..7064
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7283..7387)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7462..7481)
                     /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 ggcaaacctc gacaaatata ccgagacctt taagatggga agcaatagta cgtctactgc
      901 tgaaatctac tgcaacgtga caaacgtgaa attccagtac agcctctacg ccacaaccta
      961 cattctgatt tttattcctg gcctgttggc taatagcgca gccctttggg tgctctgtcg
     1021 gttcatctct aagaaaaaca aagctatcat attcatgatt aaccttagcg tcgccgactt
     1081 ggcccacgtg ctgtccctgc cactccgaat ctattattat ataagtcatc attggccctt
     1141 tcaacgcgcc ctgtgcctcc tgtgcttcta tctcaagtac ctgaacatgt acgcctccat
     1201 ctgctttttg acgtgcattt ctctccagag atgctttttt cttctcaagc ccttcagggc
     1261 ccgggattgg aaaaggagat acgatgtggg tatctccgct gcaatttgga tagtggtggg
     1321 taccgcatgc ctccctttcc caatactgcg cagcacggac cttaacaata acaagagttg
     1381 ctttgctgac cttgggtaca aacagatgaa tgctgtggca ctggtaggaa tgataacagt
     1441 cgcagaactc gctggattcg taatcccagt aattatcata gcttggtgca cttggaagac
     1501 cacaatcagc ctgcggcagc ctccaatggc ctttcagggt atcagcgaga ggcagaaggc
     1561 gctgcggatg gtttttatgt gcgcggccgt gtttttcatt tgttttaccc cttatcacat
     1621 caactttata ttttacacca tggtgaagga aactatcatc tcttcatgtc cagtagtgcg
     1681 gatagcactc tatttccatc ctttctgcct ctgtctggcc tcactgtgct gcctcctcga
     1741 ccccatactg tactatttca tggctagcga gttccgcgac cagctctcaa ggcacggctc
     1801 atctgtgacg agaagccgcc tgatgtccaa ggagtccggc tcttcaatga ttgggatcga
     1861 taccggtgga cgcaccccac ccagcctggg tccccaagat gagtcctgca ccaccgccag
     1921 ctcctccctg gccaaggaca cttcatcgac cggtgagaac ctgtacttcc agctaagatt
     1981 agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg
     2041 tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca
     2101 tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca
     2161 tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa
     2221 aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg
     2281 gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg
     2341 tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta ctttaggttg
     2401 cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga
     2461 tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc
     2521 agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga
     2581 aagtgggtcc gcgtacagcc gcgcgcgtac gaaaaacaat tacgggtcta ccatcgaggg
     2641 cctgctcgat ctcccggacg acgacgcccc cgaagaggcg gggctggcgg ctccgcgcct
     2701 gtcctttctc cccgcgggac acacgcgcag actgtcgacg gcccccccga ccgatgtcag
     2761 cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct
     2821 agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc
     2881 ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt
     2941 taccgatgcc cttggaattg acgagtacgg tgggtagtct agagggcccg tttaaacccg
     3001 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3061 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3121 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3181 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3241 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3301 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3361 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3421 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3481 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3541 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3601 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3661 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3721 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3781 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3901 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3961 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4021 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4081 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4141 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4201 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4261 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4321 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4381 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4441 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4501 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4561 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4621 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4681 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4741 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4801 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4861 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4921 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4981 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5041 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5221 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5281 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5341 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5401 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5461 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5521 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5581 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5641 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5701 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5761 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5821 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5881 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5941 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6001 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6061 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6121 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6181 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6241 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6301 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6361 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6421 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6481 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6541 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6601 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6661 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6721 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6781 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6841 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6901 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6961 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7021 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7081 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7141 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7201 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7261 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7321 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7381 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7441 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     7501 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     7561 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     7621 gcttcgcgat gtacgggcca gatatacgcg tt
//
