LOCUS       sequence-173899-        7763 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 7763)
  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 7763)
  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: GPER-Tango
FEATURES             Location/Qualifiers
     source          1..7763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2318..2932,2933..3319)
                     /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(2385..2404)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3357..3374)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3363..3587
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3633..4061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3720..3739)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3930..3951
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4070..4090)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4075..4404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4255..4390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4317..4336
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4471..5265
                     /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(4525..4544)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5135..5154
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5439..5560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5476..5495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5530..5549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5609..5625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5609..5625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5622..5644)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5633..5649
                     /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(5657..5663,5664..5681,5682..5687))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5702..5723
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5840..5857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6011..6596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6091..6110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6767..7558,7559..7627))
                     /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     7390..7409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7628..7732)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgcgccac catgaagacg atcatcgccc tgagctacat
     1021 cttctgcctg gtattcgccg actacaagga cgatgatgac gccagcatcg atatggacgt
     1081 aacatcccaa gcaagaggag tgggtcttga gatgtacccg gggactgcgc agcccgcagc
     1141 tcctaacact acttcacccg agctgaacct gtcacaccct ctcctgggca ctgcgctcgc
     1201 gaacggaacc ggggagctgt ctgagcacca gcagtacgtc ataggcctgt tcctgtcctg
     1261 tttgtatacc atttttctct tccccattgg gtttgtgggg aacatcctga ttcttgttgt
     1321 taacatcagc ttcagggaga aaatgacgat tcctgacttg tactttatca acctggccgt
     1381 cgccgacttg attttggtgg cggattctct gattgaggtt tttaacctcc acgagagata
     1441 ttacgacatc gctgtgctgt gtacctttat gagcctcttt ctgcaggtta acatgtattc
     1501 ctcagtgttt tttctgacct ggatgtcatt cgacagatac atcgccctcg ccagggctat
     1561 gaggtgctct ctgttccgca cgaaacacca cgcccggctt agttgcgggc tcatttggat
     1621 ggcctcagtg agcgcaacac tggtgccgtt tactgccgta cacctccagc atacagacga
     1681 agcctgtttt tgcttcgctg atgttaggga ggtccaatgg ctggaagtta ctctgggttt
     1741 catagttcca tttgccatca tagggctttg ctatagcttg atcgttcgag tactggtgag
     1801 agcacacaga caccgcggac tcaggcctcg ccgccagaag gcactgagga tgatcctcgc
     1861 cgtcgtgctg gtattttttg tgtgctggct gccagagaat gtcttcatta gcgttcattt
     1921 gctgcagagg actcaacctg gtgccgcccc ctgtaagcag agcttcaggc acgcccaccc
     1981 cctcacaggc catatcgtga atttggccgc tttttctaat tcctgtctga accctctgat
     2041 ctacagtttc ctgggggaaa cctttaggga caaactgcga ttgtacatag aacaaaagac
     2101 caacctgccg gccctgaatc ggttctgcca cgcggccctc aaagctgtga ttccggactc
     2161 aactgagcag agtgatgtcc gatttagctc tgcggtcatc gataccggtg gacgcacccc
     2221 acccagcctg ggtccccaag atgagtcctg caccaccgcc agctcctccc tggccaagga
     2281 cacttcatcg accggtgaga acctgtactt ccagctaaga ttagataaaa gtaaagtgat
     2341 taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact
     2401 cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc
     2461 tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt
     2521 agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt
     2581 actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta
     2641 tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc
     2701 attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga
     2761 gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt
     2821 acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct
     2881 tgaattgatc atatgcggat tagaaaaaca acttaaatgt gaaagtgggt ccgcgtacag
     2941 ccgcgcgcgt acgaaaaaca attacgggtc taccatcgag ggcctgctcg atctcccgga
     3001 cgacgacgcc cccgaagagg cggggctggc ggctccgcgc ctgtcctttc tccccgcggg
     3061 acacacgcgc agactgtcga cggccccccc gaccgatgtc agcctggggg acgagctcca
     3121 cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga
     3181 catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta
     3241 cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat
     3301 tgacgagtac ggtgggtagc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3361 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3421 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3481 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3541 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3601 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3661 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3721 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3781 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3841 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3901 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3961 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4021 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4081 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4141 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4201 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4261 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4321 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4381 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4441 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4501 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4561 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4621 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4681 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4741 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4801 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4861 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4921 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4981 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5041 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5101 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5161 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5221 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5281 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5341 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5401 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5461 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5521 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5581 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5641 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5701 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5821 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6541 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6601 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6661 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6721 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6841 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6961 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7201 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7261 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7441 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7621 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7681 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7741 cccgaaaagt gccacctgac gtc
//
