LOCUS       sequence-456562-        7994 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 7994)
  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 7994)
  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: GPR39-Tango
FEATURES             Location/Qualifiers
     source          1..7994
                     /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(2552..3166,3167..3553)
                     /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(2619..2638)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3585..3602)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3591..3815
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3861..4289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3948..3967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4158..4179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4298..4318)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4303..4632
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4483..4618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4545..4564
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4699..5493
                     /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(4753..4772)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5363..5382
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5667..5788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5704..5723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5758..5777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5837..5853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5837..5853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5850..5872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5861..5877
                     /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(5885..5891,5892..5909,5910..5915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5930..5951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6068..6085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6239..6827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6319..6338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6998..7789,7790..7858))
                     /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     7621..7640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7859..7963)
                     /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 atatggcctc
     1081 accttcactg cccggttccg actgtagcca gattattgac catagccacg tgccggagtt
     1141 tgaagtggcc acatggataa aaattaccct gatcctggtg tatctcatca tattcgtgat
     1201 ggggttgctt gggaattctg ccactattag agtcacccaa gtgctgcaaa aaaagggcta
     1261 cctgcagaaa gaagtgaccg atcatatggt gagcctcgcc tgttcagaca ttctcgtctt
     1321 cctgatcggt atgcccatgg aattctacag tattatttgg aatcccctga ctacatcaag
     1381 ctacacactt tcttgcaagt tgcatacttt cctgttcgaa gcgtgcagtt acgccaccct
     1441 gctgcacgtg ctgacactct catttgagag atacatcgca atctgtcatc cgttcaggta
     1501 caaagccgta agtggccctt gtcaggtcaa gctgctgatt gggtttgtgt gggtgaccag
     1561 cgccttggtg gccttgccac ttctttttgc tatgggaaca gagtaccctc tcgtcaacgt
     1621 cccaagtcac cgaggcctca cgtgtaatag gtcaagtaca cgacatcacg aacagccaga
     1681 gacttccaat atgagtatct gcactaatct gtcctcccgc tggaccgttt tccagtcttc
     1741 aatctttggc gcatttgtgg tgtacttggt cgtgctgctg tccgtagcct ttatgtgttg
     1801 gaacatgatg caggtgctca tgaagagcca gaaaggcagc ttggccgggg gtacccggcc
     1861 gcctcagctt cgcaaatcag aaagcgagga gtcccggact gcccgccggc agactattat
     1921 ttttctgcga ctgatagtgg tgacccttgc cgtgtgttgg atgcccaatc agatacgcag
     1981 aatcatggca gcagctaaac ccaaacatga ttggacccgg tcctacttcc gggcctacat
     2041 gattctgctg ccgttcagtg agacgttctt ctacctgagc agtgtcatta atccacttct
     2101 ttacactgtg agcagtcagc aattcaggag ggtcttcgtc caagtcttgt gctgcagact
     2161 cagcctccag cacgcgaacc acgaaaaaag gctccgggta cacgcgcact ccacaacgga
     2221 ctccgcacga ttcgttcaga gaccccttct ttttgcttca cgaagacaat ccagtgccag
     2281 acggacggaa aagatattcc tgtctacttt tcaatcagaa gccgagcccc agagcaagag
     2341 ccagtccctg tccttggaaa gcctggagcc gaatagcggc gcaaaacctg ctaactctgc
     2401 tgcggagaat gggtttcaag aacacgaggt gatcgatacc ggtggacgca ccccacccag
     2461 cctgggtccc caagatgagt cctgcaccac cgccagctcc tccctggcca aggacacttc
     2521 atcgaccggt gagaacctgt acttccagct aagattagat aaaagtaaag tgattaacag
     2581 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
     2641 gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct
     2701 cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg
     2761 ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag
     2821 tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac
     2881 tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata
     2941 tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca
     3001 agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca
     3061 agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt
     3121 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagccgcgc
     3181 gcgtacgaaa aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga
     3241 cgcccccgaa gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac
     3301 gcgcagactg tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga
     3361 cggcgaggac gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt
     3421 gggggacggg gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc
     3481 tctggatatg gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga
     3541 gtacggtggg tagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3601 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3661 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3721 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3781 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3841 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3901 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3961 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4021 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4081 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4141 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4201 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4261 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4321 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4381 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4441 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4501 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4561 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4621 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4681 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4741 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4801 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4861 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4921 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4981 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5041 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5101 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5161 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5221 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5281 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5341 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5401 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5461 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5521 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5581 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5641 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5701 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5761 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5821 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5881 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5941 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6001 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6061 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6901 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6961 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7021 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7081 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7141 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7201 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7261 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7321 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7381 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7441 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7501 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7561 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7621 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7681 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7741 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7801 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7861 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7921 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7981 tgccacctga cgtc
//
