LOCUS       sequence-263503-        7694 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 7694)
  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 7694)
  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: FPR3-Tango
FEATURES             Location/Qualifiers
     source          1..7694
                     /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(2252..2866,2867..3253)
                     /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(2319..2338)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3285..3302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3291..3515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3561..3989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3648..3667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3858..3879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3998..4018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4003..4332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4183..4318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4245..4264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4399..5193
                     /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(4453..4472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5063..5082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5367..5488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5404..5423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5458..5477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5537..5553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5537..5553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5550..5572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5561..5577
                     /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(5585..5591,5592..5609,5610..5615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5630..5651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5768..5785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5939..6527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6019..6038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6698..7489,7490..7558))
                     /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     7321..7340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7559..7663)
                     /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 atatggagac
     1081 caacttttca atacccttga atgagacaga ggaggtcctg cctgaacccg cgggccacac
     1141 tgtgctttgg attttttcac ttctcgtcca cggtgtgact ttcgtgttcg gtgtccttgg
     1201 caacggcctc gtgatctggg ttgcaggctt cagaatgaca aggacagtga ataccatttg
     1261 ttacttgaat ttggccctgg cagattttag tttctctgcg attctgccat ttcgcatggt
     1321 gtctgtagca atgcgcgaga agtggccatt tggctcattc ctgtgtaagc ttgtgcatgt
     1381 catgattgat attaatcttt tcgtgagcgt gtatctgatt acgataatcg cactcgaccg
     1441 atgtatctgt gtgctccacc ccgcctgggc ccaaaatcat aggaccatgt ctctcgccaa
     1501 acgcgttatg actggtcttt ggatctttac cattgtattg actctgccca attttatatt
     1561 ctggaccacc atctctacca cgaatgggga tacatactgt atttttaatt tcgccttctg
     1621 gggcgataca gccgtggaaa gacttaacgt gttcatcacc atggccaaag tgtttctgat
     1681 tctccacttt atcatcggct tttccgtgcc catgtccatc atcacagtgt gctatggcat
     1741 catcgctgcc aagattcaca gaaatcatat gatcaaatca agtaggcccc tccgagtctt
     1801 cgcggccgtc gtggctagct tttttatatg ttggttcccc tatgaactga taggtatcct
     1861 gatggcagtg tggctgaagg agatgctcct gaacggtaag tataagatca tccttgtgct
     1921 gatcaatcct acatccagcc tggccttttt caacagttgt ctcaacccca tcctctacgt
     1981 gttcatgggg cgcaacttcc aggagcgcct gattcggtcc ctccctacct cattggagcg
     2041 cgccctcact gaagtccccg attctgccca aacaagtaac acggacacca catctgccag
     2101 tcctccagag gaaaccgaac tgcaggctat gatcgatacc ggtggacgca ccccacccag
     2161 cctgggtccc caagatgagt cctgcaccac cgccagctcc tccctggcca aggacacttc
     2221 atcgaccggt gagaacctgt acttccagct aagattagat aaaagtaaag tgattaacag
     2281 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
     2341 gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct
     2401 cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg
     2461 ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag
     2521 tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac
     2581 tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata
     2641 tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca
     2701 agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca
     2761 agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt
     2821 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagccgcgc
     2881 gcgtacgaaa aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga
     2941 cgcccccgaa gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac
     3001 gcgcagactg tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga
     3061 cggcgaggac gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt
     3121 gggggacggg gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc
     3181 tctggatatg gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga
     3241 gtacggtggg tagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3301 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3361 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3421 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3481 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3541 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3601 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3661 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3721 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3781 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3841 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3901 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3961 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4141 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4201 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4261 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4321 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4381 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4441 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4501 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4561 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4621 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4681 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4741 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4801 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4861 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4921 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4981 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5041 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5101 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5161 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5221 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5281 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5341 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5401 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5461 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5521 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5581 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5641 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5701 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5761 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5821 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6121 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6361 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6421 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7561 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7621 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7681 tgccacctga cgtc
//
