LOCUS       sequence-347878-        7781 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 7781)
  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 7781)
  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: S1PR1-Tango
FEATURES             Location/Qualifiers
     source          1..7781
                     /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(2105..2719,2720..3106)
                     /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(2172..2191)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3138..3155)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3144..3368
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3414..3842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3501..3520)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3711..3732
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3851..3871)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3856..4185
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4036..4171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4098..4117
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4252..5046
                     /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(4306..4325)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4916..4935
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5220..5341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5257..5276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5311..5330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5390..5406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5390..5406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5403..5425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5414..5430
                     /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(5438..5444,5445..5462,5463..5468))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5483..5504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5621..5638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5792..6380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5872..5891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6551..7342,7343..7411))
                     /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     7174..7193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7412..7516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7591..7610)
                     /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 gggaccaaca tcagtcccat tggttaaagc ccatcgaagc tccgtgagcg actacgtgaa
      901 ctacgacatc atcgtgaggc actataacta taccggaaag ctcaacatat ccgccgataa
      961 agagaatagt attaaactga cctctgtcgt cttcatactg atctgttgtt tcatcattct
     1021 cgaaaacatt ttcgtgctcc ttaccatctg gaaaaccaaa aagttccaca gacctatgta
     1081 ctattttatc ggaaatctgg ccctgtctga cttgttggcc ggggtggcct atactgcaaa
     1141 cttgctgctc tcaggcgcca ctacatacaa actcacgccc gctcagtggt ttctgaggga
     1201 gggctccatg tttgtggcgc tgagcgctag cgtgttcagc ctgctggcta tagccatcga
     1261 acgatacatt actatgctga agatgaagct gcacaatggc tctaacaatt ttaggctctt
     1321 tctccttata tccgcgtgct gggtgatttc tctgattctc ggggggctgc ccatcatggg
     1381 gtggaattgc atctctgctc tgtcatcatg ctccactgtg cttcccctgt atcataagca
     1441 ctatatattg ttttgtacca ctgttttcac cctcttgctg ctcagcattg tgattctcta
     1501 ctgccgcata tactccttgg tccggacaag atccaggcgc ctgacgttcc ggaaaaacat
     1561 ctctaaggcg tctagatcat ccgagaaaag cctggcgctc ctgaagaccg tcatcatcgt
     1621 gttgagcgtc tttatcgctt gctgggcccc cctgtttatc ttgctcctgc tggatgtagg
     1681 gtgtaaagtg aagacgtgcg atatcctgtt ccgggccgag tattttcttg tcctggcagt
     1741 cctgaactcc ggcactaatc ctatcattta taccctgacc aataaagaaa tgcgccgcgc
     1801 cttcatcagg attatgagct gctgtaagtg tccctccgga gactctgctg gcaaatttaa
     1861 gcggccaatc atagccggca tggagttctc cagatctaaa tccgacaata gctcacaccc
     1921 tcagaaagat gaaggcgaca atcctgaaac aattatgagt agtggtaatg tgaatagctc
     1981 aagcatcgat accggtggac gcaccccacc cagcctgggt ccccaagatg agtcctgcac
     2041 caccgccagc tcctccctgg ccaaggacac ttcatcgacc ggtgagaacc tgtacttcca
     2101 gctaagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     2161 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     2221 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     2281 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     2341 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     2401 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     2461 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
     2521 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     2581 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     2641 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     2701 taaatgtgaa agtgggtccg cgtacagccg cgcgcgtacg aaaaacaatt acgggtctac
     2761 catcgagggc ctgctcgatc tcccggacga cgacgccccc gaagaggcgg ggctggcggc
     2821 tccgcgcctg tcctttctcc ccgcgggaca cacgcgcaga ctgtcgacgg cccccccgac
     2881 cgatgtcagc ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc
     2941 cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg
     3001 atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga
     3061 gcagatgttt accgatgccc ttggaattga cgagtacggt gggtagtcta gagggcccgt
     3121 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3181 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3241 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3301 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3361 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3421 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3481 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3541 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3601 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3661 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3721 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3781 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3841 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3901 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3961 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4021 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4081 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4141 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4201 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4261 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4321 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4381 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4441 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4501 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4561 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4621 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4681 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4741 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4801 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4861 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4921 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4981 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5041 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5101 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5161 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5221 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5281 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5341 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5401 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5461 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5521 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5581 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5641 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5701 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5881 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7561 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7621 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7681 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     7741 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
