LOCUS       sequence-248122-        7766 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 7766)
  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 7766)
  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: GPR120-Tango
FEATURES             Location/Qualifiers
     source          1..7766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2326..2940,2941..3327)
                     /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(2393..2412)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3359..3376)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3365..3589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3635..4063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3722..3741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3932..3953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4072..4092)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4077..4406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4257..4392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4319..4338
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4473..5267
                     /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(4527..4546)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5137..5156
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5441..5562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5478..5497)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5532..5551
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5611..5627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5611..5627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5624..5646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5635..5651
                     /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(5659..5665,5666..5683,5684..5689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5704..5725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5842..5859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6093..6112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6772..7563,7564..7632))
                     /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     7395..7414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7633..7737)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattct
      961 gcagatatcc agcacagtgg cggccgcgcc accatgaaga cgatcatcgc cctgagctac
     1021 atcttctgcc tggtattcgc cgactacaag gacgatgatg acgccagcat cgatatgtct
     1081 cctgagtgcg cgagggctgc tggagacgca cccctgaggt cccttgagca agccaaccgc
     1141 actagattcc cctttttctc tgacgtcaag ggtgaccaca ggctggtgct tgcagccgta
     1201 gagactacgg tcctcgttct gattttcgcc gttagtctct tggggaatgt gtgtgctctg
     1261 gtcctggttg caaggcggag gcgcagaggc gccaccgcat gcctcgtgtt gaatctgttt
     1321 tgtgccgatc tcctcttcat tagcgccatt cctctcgtcc tggccgtgcg ctggaccgaa
     1381 gcttggctgc tgggccccgt ggcttgtcac ctccttttct atgttatgac cctgtctggt
     1441 tcagtgacaa tcctgaccct tgcagccgtg tccctggaga ggatggtctg catcgttcat
     1501 cttcaaagag gtgtgcgggg ccctggcagg agggcccggg ccgttctctt ggctcttatc
     1561 tggggatact ctgcagtcgc cgccctcccc ctttgtgtct tttttcgcgt cgtgccacag
     1621 cggctccctg gcgcagacca agaaattagc atttgcaccc tcatttggcc tacgatcccc
     1681 ggtgagattt catgggatgt gtccttcgtg accctgaatt ttttggtacc tggccttgtg
     1741 attgtgatct cttacagcaa aatcctccag accagcgagc acctcctgga tgccagggct
     1801 gtggtgactc actccgaaat taccaaggct agtcgcaaga ggctgaccgt gtccctcgct
     1861 tattcagaaa gtcaccagat tcgcgtgtca cagcaggact ttagactttt taggacgctg
     1921 tttttgctga tggtgagttt ctttattatg tggtccccca ttattattac aatcctgctg
     1981 attcttatcc agaattttaa acaggatctt gtgatctggc ccagcctgtt tttctgggtg
     2041 gtggcattca ctttcgctaa ctcagccctc aatcccatcc tctataatat gacgctttgc
     2101 agaaacgagt ggaagaagat cttctgctgc ttctggtttc cagagaaagg cgctatcctg
     2161 actgatacaa gtgtgaaacg aaatgacctg tctattataa gcgggatcga taccggtgga
     2221 cgcaccccac ccagcctggg tccccaagat gagtcctgca ccaccgccag ctcctccctg
     2281 gccaaggaca cttcatcgac cggtgagaac ctgtacttcc agctaagatt agataaaagt
     2341 aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc
     2401 cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat
     2461 aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt
     2521 tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga
     2581 tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa
     2641 aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta
     2701 gagaatgcat tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa
     2761 gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg
     2821 ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta
     2881 ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc
     2941 gcgtacagcc gcgcgcgtac gaaaaacaat tacgggtcta ccatcgaggg cctgctcgat
     3001 ctcccggacg acgacgcccc cgaagaggcg gggctggcgg ctccgcgcct gtcctttctc
     3061 cccgcgggac acacgcgcag actgtcgacg gcccccccga ccgatgtcag cctgggggac
     3121 gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct agacgatttc
     3181 gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc ccacgactcc
     3241 gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt taccgatgcc
     3301 cttggaattg acgagtacgg tgggtagtct agagggcccg tttaaacccg ctgatcagcc
     3361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3541 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3601 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4081 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4141 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4201 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4321 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4381 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4441 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4501 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4561 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4621 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4681 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4741 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4801 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4861 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4921 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4981 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5041 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5101 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5161 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5221 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5281 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5341 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5401 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5581 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5641 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5701 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5761 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5821 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5881 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6661 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6721 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6781 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6841 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6901 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6961 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7021 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7081 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7141 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7201 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7261 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7321 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7381 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7441 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7501 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7561 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7621 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7741 atttccccga aaagtgccac ctgacg
//
