LOCUS       sequence-115875-        8027 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 8027)
  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 8027)
  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: GLP1R-Tango
FEATURES             Location/Qualifiers
     source          1..8027
                     /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(2582..3196,3197..3583)
                     /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(2649..2668)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3621..3638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3627..3851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3897..4325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3984..4003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4194..4215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4334..4354)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4339..4668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4519..4654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4581..4600
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4735..5529
                     /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(4789..4808)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5399..5418
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5703..5824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5740..5759)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5794..5813
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5873..5889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5873..5889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5886..5908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5897..5913
                     /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(5921..5927,5928..5945,5946..5951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5966..5987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6104..6121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6275..6860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6355..6374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7031..7822,7823..7891))
                     /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     7654..7673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7892..7996)
                     /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 atatggcagg
     1081 tgcccctgga cccctcaggc tcgcactgct gcttctgggg atggtcggac gagccggacc
     1141 cagacctcag ggcgctaccg tgagtctttg ggagaccgtc cagaagtggc gggagtatcg
     1201 ccgccagtgt caacgcagcc tgacagaaga tcctcctcca gccaccgact tgttttgcaa
     1261 taggacattt gatgagtatg cctgctggcc agacggcgag cccggctcat tcgtgaatgt
     1321 gtcatgtcca tggtacctgc catgggcatc atctgtgcct caaggacacg tctacaggtt
     1381 ctgcacagct gagggcctct ggctgcaaaa agataacagt tcccttccat ggcgcgatct
     1441 tagcgagtgc gaggaatcta agaggggcga gcgcagttcc cccgaggaac agctgctgtt
     1501 tctgtacatt atctacaccg tcggctacgc gctgtccttt tccgctctgg ttatcgctag
     1561 tgccatcctt ttggggtttc gccaccttca ctgcacaaga aattacatcc acttgaatct
     1621 ctttgcaagt tttattctgc gggccttgag tgtgttcatt aaggacgctg ccctgaagtg
     1681 gatgtactct accgcggccc aacagcacca atgggatgga ctcctcagct atcaggattc
     1741 tctttcctgt cgactcgtgt ttctgcttat gcagtactgt gtggcagcca attattattg
     1801 gctgctggtc gaaggagtat atctttacac actgctcgca ttctccgtgc tgagtgaaca
     1861 gtggattttc aggctttacg tgtccattgg atggggtgtg cctctgctgt tcgtggtgcc
     1921 ttggggcatc gtcaagtacc tgtatgaaga tgaaggctgt tggaccagaa actctaacat
     1981 gaactactgg ctcatcatac gcctgcccat actcttcgcg attggagtca actttcttat
     2041 atttgtgaga gtgatctgta tcgtcgtgtc aaagctgaaa gctaacctta tgtgcaaaac
     2101 cgacattaag tgcagactgg ctaagtccac tctcaccctc atcccgctgc tcggaacaca
     2161 tgaagtgata tttgccttcg tgatggacga gcatgcgaga ggcacactgc ggtttatcaa
     2221 actgttcaca gagctgtctt tcactagttt ccaaggattg atggtggcta ttctctactg
     2281 cttcgtgaac aatgaggtcc aacttgaatt tcggaagtca tgggaacggt ggcggcttga
     2341 acacctccat attcagcgcg atagttctat gaagccactt aaatgtccca cttccagcct
     2401 gagttccgga gccaccgccg gctcttcaat gtatacagcc acttgtcagg cgtcctgctc
     2461 tatcgatacc ggtggacgca ccccacccag cctgggtccc caagatgagt cctgcaccac
     2521 cgccagctcc tccctggcca aggacacttc atcgaccggt gagaacctgt acttccagct
     2581 aagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
     2641 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
     2701 ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag
     2761 gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa
     2821 cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg
     2881 tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca
     2941 acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt
     3001 aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac
     3061 tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc
     3121 agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa
     3181 atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa aacaattacg ggtctaccat
     3241 cgagggcctg ctcgatctcc cggacgacga cgcccccgaa gaggcggggc tggcggctcc
     3301 gcgcctgtcc tttctccccg cgggacacac gcgcagactg tcgacggccc ccccgaccga
     3361 tgtcagcctg ggggacgagc tccacttaga cggcgaggac gtggcgatgg cgcatgccga
     3421 cgcgctagac gatttcgatc tggacatgtt gggggacggg gattccccgg gtccgggatt
     3481 taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca
     3541 gatgtttacc gatgcccttg gaattgacga gtacggtggg tagctcgagt ctagagggcc
     3601 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3661 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3721 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3781 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3841 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3901 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3961 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4021 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4081 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4141 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4201 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4261 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4321 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4381 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4501 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4561 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4621 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4681 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4741 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4801 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4861 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4921 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4981 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5041 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5101 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5161 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5221 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5281 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5341 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5401 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5461 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5521 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5581 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5641 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5701 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5761 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5821 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5881 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5941 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6001 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6061 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6121 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6181 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6241 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6781 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7021 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7081 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7141 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7201 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7261 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7321 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7381 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7441 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7501 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7561 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7621 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7681 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7741 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7801 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7861 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7921 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7981 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
