LOCUS       sequence-456481-        8315 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 8315)
  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 8315)
  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: GPR123-Tango
FEATURES             Location/Qualifiers
     source          1..8315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     264..283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        358..462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(463..531,532..1323)
                     /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     complement(681..700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1494..2082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1983..2002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2236..2253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2370..2391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2406..2411,2412..2429,2430..2436)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2444..2460
                     /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)."
     primer_bind     2449..2471
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2468..2484
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2468..2484
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2533..2654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2544..2563)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2598..2617
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2828..3622)
                     /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(2939..2958)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3549..3568
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3689..4018)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3703..3838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3757..3776)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4003..4023
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4032..4460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4142..4163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4354..4373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4506..4730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4719..4736
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(4768..5154,5155..5769))
                     /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     5683..5702
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7760..7779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7761..7779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7824..8027)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7853..7873)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(8028..8315,1..92)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
       61 ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg
      121 aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg
      181 ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg
      241 gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag
      301 gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc
      361 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      421 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      481 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      541 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      601 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      661 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
      721 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      781 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      841 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      901 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      961 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1021 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1081 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1141 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1201 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1261 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1321 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1381 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1441 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1561 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1621 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1681 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1741 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1801 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1861 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1921 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1981 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2041 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2101 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2161 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2221 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2281 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2341 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2401 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2461 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgacggt
     2521 atacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2581 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2641 gcaataaaca agttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     2701 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     2761 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     2821 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     2881 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     2941 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     3001 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     3061 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     3121 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     3181 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     3241 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     3301 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     3361 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     3421 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     3481 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     3541 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     3601 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatccga
     3661 aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc
     3721 ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa
     3781 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     3841 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     3901 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     3961 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     4021 tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4081 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4141 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4201 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4261 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4321 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4381 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4441 tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc
     4501 agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc
     4561 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     4621 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     4681 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga
     4741 tcagcgggtt taaacgggcc ctctagacta cccaccgtac tcgtcaattc caagggcatc
     4801 ggtaaacatc tgctcaaact cgaagtcggc catatccaga gcgccgtagg gggcggagtc
     4861 gtggggggta aatcccggac ccggggaatc cccgtccccc aacatgtcca gatcgaaatc
     4921 gtctagcgcg tcggcatgcg ccatcgccac gtcctcgccg tctaagtgga gctcgtcccc
     4981 caggctgaca tcggtcgggg gggccgtcga cagtctgcgc gtgtgtcccg cggggagaaa
     5041 ggacaggcgc ggagccgcca gccccgcctc ttcgggggcg tcgtcgtccg ggagatcgag
     5101 caggccctcg atggtagacc cgtaattgtt tttcgtacgc gcgcggctgt acgcggaccc
     5161 actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc cgaataagaa
     5221 ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata atggcggcat
     5281 actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt gatcttccaa
     5341 tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat tctctagtga
     5401 aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact gtttttctgt
     5461 aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag cacatctaaa
     5521 acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag ggcaaaagtg
     5581 agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc gcttattttt
     5641 tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt tacgggttgt
     5701 taaaccttcg attccgacct cattaagcag ctctaatgcg ctgttaatca ctttactttt
     5761 atctaatctt agctggaagt acaggttctc accggtcgat gaagtgtcct tggccaggga
     5821 ggagctggcg gtggtgcagg actcatcttg gggacccagg ctgggtgggg tgcgtccacc
     5881 ggtatcgatc actgtggtct cgttcttcca tggtccggtt cgtatattac ccgtgccgtc
     5941 tgtgccgaac ggcagaccct ccaggagttt tcctttaggc agcccattct gggaaggtcc
     6001 gccaaagggc aaggactgag ttcgtctaag catttccaag tgaccaggac caagtgcggc
     6061 ctctctccca ggctgagttg gacatgagta caatgctgga gacccatcgc ttccttcggg
     6121 ctgtgtgacc atagggaatg ggtcgccctg ggtgcaacag gcaagagtgt gagtccctgc
     6181 tgggccatcc agggaagatg gtgggctatc tgttcgggat gagcgtgggg agccatcgag
     6241 ggagctaggg atgtggtagg catattcggg ctcttcggca ggatggcgtg aaaaatacgg
     6301 tggctttgtc cgtccttgca ggcagccatg cagatggggg tcgtggccga acgcaccttc
     6361 ttcctgcaag tgcacgtggg cttcgtcggc ggtcagtggc tcgcagtgca tcttggcgca
     6421 acagggggta gctgggctca gacaactggc atgcccctga gcggcctgca ggttagtcat
     6481 tttacagggg ccgggggggt gagcaaagcc gcggggctgc cccagaccag gtgagtgaag
     6541 acaggctgct cttccaagag cggctccgtt agcgtccaga gctgggtggg cgtctttcct
     6601 tggagggcag caggcccacc aacactgcca gacgtcttcc cgtttagcac aatggtggat
     6661 caacacaaag agtccgagtg tcacgcaaaa ggcaccgtac agacaggaga aaaccatatc
     6721 cagaaaatgt ccttgagaca ctgcgagagc cccaaatgcc catgtggcgg taaagaggaa
     6781 gagtgtaaag gcggcagctc tcagttgagc ttgaaagctg tgctcgttct gcaggacaga
     6841 tgctccggga gcatcgtgtg cgggaggagt cccagggcga atacccctcc cgccctcggg
     6901 tgtagccagc ctgcgttgct cttcaggctg tgtcctaagc tcataccttc tgccggggtg
     6961 tctcctcagc tgcacatatg tgcccagaaa atacacgcag gtcaccaaag tgattatggc
     7021 tgctggtccg taaaaggctc ccaaagaagg ttcccaggcc atccagcagt aggctgtgtc
     7081 ctcgtcctcg gtgccataat tcctaatgtt tgtggcggct gtcacgccac aaattatgaa
     7141 gggcaccccg ccacttacca aatagaagcg gaggaggggc tgcctcggat agggaggctg
     7201 atcagtgtcc aggcacagtg gtgctttctt agttacctgt ttgtatatat tgcgggctgt
     7261 gactccgatc cagagcatgg tgctcaatgt tgagtaatgc agcacgatgc ccacggcttg
     7321 acacagtatt gggtatttcg tccgattgat gccaccagca aagaccgtaa aagtcaaggc
     7381 cgcgtggaaa cagaagttca gcagagtgtg tcggcccttg cgagagattc gtatggcact
     7441 ttggtgtacg atataagtca caaaggaggc gagcagacac aacagcatga cggcagtgca
     7501 cgcgtagacc acggggtgaa ggaattcgcc tgggtatctc ggaagggaga gcacggtttt
     7561 cagatccata tcgatgctgg cgtcatcatc gtccttgtag tcggcgaata ccaggcagaa
     7621 gatgtagctc agggcgatga tcgtcttcat ggtggcgcgg ccgccactgt gctggatatc
     7681 tgcagaattc caccacactg gactagtgga tccgagctcg gtaccaagct taagtttaaa
     7741 cgctagccag cttgggtctc cctatagtga gtcgtattaa tttcgataag ccagtaagca
     7801 gtgggttctc tagttagcca gagagctctg cttatataga cctcccaccg tacacgccta
     7861 ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt
     7921 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     7981 tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg
     8041 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     8101 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     8161 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     8221 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     8281 gggtcgttgg gcggtcagcc aggcgggcca tttac
//
