LOCUS       sequence-344663-        7892 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 7892)
  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 7892)
  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: GPR151-Tango
FEATURES             Location/Qualifiers
     source          1..7892
                     /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(2216..2830,2831..3217)
                     /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(2283..2302)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3249..3266)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3255..3479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3525..3953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3612..3631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3822..3843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3962..3982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3967..4296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4147..4282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4209..4228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4363..5157
                     /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(4417..4436)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5027..5046
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5331..5452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5368..5387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5422..5441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5501..5517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5501..5517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5514..5536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5525..5541
                     /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(5549..5555,5556..5573,5574..5579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5594..5615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5903..6491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5983..6002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6662..7453,7454..7522))
                     /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     7285..7304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7523..7627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7702..7721)
                     /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 gttggccgcc gcatttgccg attcaaatag ttccagtatg aacgtttcct tcgctcacct
      901 tcattttgct ggtggctact tgcccagcga cagccaagat tggcggacaa ttattcccgc
      961 tctcctggtt gcagtatgtt tggtgggttt tgtggggaat ctttgcgtca tcggtatact
     1021 tctgcacaac gcctggaaag gcaagccatc catgatccac agcctcattc ttaatctgtc
     1081 tctggcagac ctctccctct tgctgttcag cgccccgata cgggcgacag cgtacagtaa
     1141 gagtgtctgg gacttgggct ggtttgtatg caaaagttcc gattggttta tccacacctg
     1201 tatggccgca aagtccctga caatcgtggt cgtggcaaag gtttgcttta tgtatgccag
     1261 tgaccctgct aagcaagtgt ctattcataa ttatactata tggtcagtgc tggtggccat
     1321 ctggacagtg gcctctctcc tgcccctgcc agaatggttt ttttccacca tccgccacca
     1381 cgagggggtt gagatgtgct tggtggacgt gccggccgtg gctgaggagt tcatgtctat
     1441 gtttggaaag ttgtacccat tgcttgcatt cgggctgccg ctctttttcg catcattcta
     1501 cttctggaga gcatatgatc agtgcaaaaa aagaggtacc aagacacaga atctgaggaa
     1561 ccaaattagg agtaaacagg tgacagtgat gcttctctct attgccatca tctccgcgct
     1621 gctgtggctg ccagaatggg tggcttggct ctgggtatgg catctcaaag cagccggccc
     1681 tgccccacct cagggcttta tcgcactctc ccaggtcctg atgttctcta tctccagtgc
     1741 caatcccctg atcttcctcg tgatgtccga agaatttaga gagggtctca aaggagtctg
     1801 gaagtggatg ataacaaaaa agcccccaac agtgagcgag tcacaggaga caccggccgg
     1861 aaatagtgaa ggtcttccgg ataaggtacc tagtcccgaa agccctgcct ccatcccgga
     1921 gaaggaaaaa ccgagctccc cctcctccgg aaaaggaaag acagaaaagg cagagatacc
     1981 gattcttcca gacgtagagc aattctggca cgagcgcgac accgtgccca gcgtgcagga
     2041 taatgatcca atcccttggg agcatgagga ccaagaaact ggagaaggag tgaagatcga
     2101 taccggtgga cgcaccccac ccagcctggg tccccaagat gagtcctgca ccaccgccag
     2161 ctcctccctg gccaaggaca cttcatcgac cggtgagaac ctgtacttcc agctaagatt
     2221 agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg
     2281 tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca
     2341 tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca
     2401 tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa
     2461 aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg
     2521 gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg
     2581 tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta ctttaggttg
     2641 cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga
     2701 tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc
     2761 agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga
     2821 aagtgggtcc gcgtacagcc gcgcgcgtac gaaaaacaat tacgggtcta ccatcgaggg
     2881 cctgctcgat ctcccggacg acgacgcccc cgaagaggcg gggctggcgg ctccgcgcct
     2941 gtcctttctc cccgcgggac acacgcgcag actgtcgacg gcccccccga ccgatgtcag
     3001 cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct
     3061 agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc
     3121 ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt
     3181 taccgatgcc cttggaattg acgagtacgg tgggtagtct agagggcccg tttaaacccg
     3241 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3301 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3361 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3421 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3481 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3601 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3721 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3841 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3961 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4201 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4261 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4321 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4381 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4441 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4501 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4561 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4621 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4681 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4741 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4801 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4861 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4921 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4981 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5041 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5101 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5161 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5221 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5281 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5461 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5521 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5581 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5641 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5701 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5761 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5821 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5881 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5941 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6001 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6061 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6121 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6181 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6241 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6301 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6361 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6421 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6481 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6541 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6601 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6661 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6721 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6781 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6841 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6901 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6961 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7021 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7081 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7141 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7201 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7261 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7321 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7381 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7441 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7501 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7561 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7621 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7681 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     7741 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     7801 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     7861 gcttcgcgat gtacgggcca gatatacgcg tt
//
