LOCUS       sequence-251911-        8072 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 8072)
  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 8072)
  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: HTR7-Tango
FEATURES             Location/Qualifiers
     source          1..8072
                     /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(2632..3246,3247..3633)
                     /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(2699..2718)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3665..3682)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3671..3895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3941..4369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4238..4259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4378..4398)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4383..4712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4563..4698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4625..4644
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4779..5573
                     /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(4833..4852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5443..5462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5747..5868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5784..5803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5838..5857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5917..5933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5917..5933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5930..5952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5941..5957
                     /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(5965..5971,5972..5989,5990..5995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6010..6031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6148..6165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6319..6907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6399..6418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7078..7869,7870..7938))
                     /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     7701..7720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7939..8043)
                     /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 cgatatgatg
     1081 gacgttaaca gctctggaag acctgatctg tatggccacc tgcggagctt tctcttgccc
     1141 gaagtcggca gaggcctgcc tgacctcagc cctgatggag gcgccgaccc cgtcgcagga
     1201 tcctgggccc cacatctgct gtccgaagta acggcgtccc ctgctccaac ctgggatgcc
     1261 ccccctgata atgcctccgg gtgcggcgaa cagataaatt atggacgagt ggaaaaggta
     1321 gtgatagggt caatactgac tcttattact cttctgacta tcgctggcaa ttgcctcgta
     1381 gtaatctcag tttgttttgt aaaaaaactc cggcagccca gtaactacct gatcgtctca
     1441 ctcgcactgg ccgatctcag tgtggcagtc gctgttatgc cgttcgtctc tgtcacagat
     1501 ctgataggtg gtaagtggat cttcggacat tttttttgta acgtgtttat tgcgatggac
     1561 gttatgtgtt gcaccgcctc aattatgact ctctgtgtga tcagtatcga cagatacttg
     1621 ggcattacac gccccttgac atacccagtg agacaaaatg gaaagtgcat ggccaagatg
     1681 attctcagtg tatggctgct tagcgccagc atcacccttc ctccactctt tgggtgggca
     1741 caaaatgtaa acgatgataa agtctgtttg atttcacaag actttggtta tacgatttat
     1801 tccaccgctg tcgcctttta cattcccatg tctgtgatgc tgttcatgta ctatcagatc
     1861 tataaggcgg ctcgaaagag tgccgccaaa cataagtttc cggggttccc cagagtggag
     1921 cccgatagcg taatcgcact gaacggtatt gtgaaactgc agaaagaagt ggaagagtgt
     1981 gcgaatctct ctagactgct taagcatgag cgaaaaaata tctccatctt caagagggaa
     2041 caaaaagccg ccaccaccct cggtatcatc gtgggagcgt tcacggtgtg ttggctgcct
     2101 ttcttcctcc tgtccactgc tagaccattc atttgcggaa cttcctgctc ttgtattccg
     2161 ctgtgggttg aaaggacctt tctgtggctg ggttacgcta actcactgat taatccattc
     2221 atttacgcat ttttcaaccg ggacctgagg accacctaca gatccttgtt gcagtgccag
     2281 taccggaaca taaaccgcaa actttcagcc gccggaatgc acgaagccct gaaactggcc
     2341 gaaagacctg aaaggcccga attcgtcctt agagcctgta ctcgccgcgt cttgcttagg
     2401 cccgagaaga ggcctcccgt gagtgtgtgg gtcttgcaga gcccagatca tcacaactgg
     2461 ctggccgaca agatgcttac tacagttgag aagaaggtaa tgatccatga catcgatacc
     2521 ggtggacgca ccccacccag cctgggtccc caagatgagt cctgcaccac cgccagctcc
     2581 tccctggcca aggacacttc atcgaccggt gagaacctgt acttccagct aagattagat
     2641 aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta
     2701 acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta
     2761 aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact
     2821 cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt
     2881 tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct
     2941 acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt
     3001 tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta
     3061 ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt
     3121 atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc
     3181 ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt
     3241 gggtccgcgt acagccgcgc gcgtacgaaa aacaattacg ggtctaccat cgagggcctg
     3301 ctcgatctcc cggacgacga cgcccccgaa gaggcggggc tggcggctcc gcgcctgtcc
     3361 tttctccccg cgggacacac gcgcagactg tcgacggccc ccccgaccga tgtcagcctg
     3421 ggggacgagc tccacttaga cggcgaggac gtggcgatgg cgcatgccga cgcgctagac
     3481 gatttcgatc tggacatgtt gggggacggg gattccccgg gtccgggatt taccccccac
     3541 gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca gatgtttacc
     3601 gatgcccttg gaattgacga gtacggtggg tagtctagag ggcccgttta aacccgctga
     3661 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3721 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3781 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3841 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3901 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3961 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4021 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4081 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4141 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4201 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4261 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4321 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4381 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4441 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4501 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4561 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4621 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4681 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4741 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4801 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4861 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4921 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4981 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5041 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5101 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5161 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5221 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5281 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5341 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5401 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5461 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5521 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5581 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5641 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5701 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5761 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5821 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5881 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5941 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6001 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6061 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6121 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6721 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacctga cg
//
