LOCUS       sequence-456477-        8066 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 8066)
  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 8066)
  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: ADRB1-Tango
FEATURES             Location/Qualifiers
     source          1..8066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1269..1288)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1270..1288)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1333..1536)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1362..1382)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1537..1916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2088..2107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2182..2286
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2287..2355,2356..3147)
                     /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(2505..2524)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3318..3906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3807..3826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4060..4077
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4194..4215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4230..4235,4236..4253,4254..4260)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4268..4284
                     /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     4273..4295
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4292..4308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4292..4308
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4357..4478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4368..4387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4422..4441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4652..5446)
                     /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(4763..4782)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5373..5392
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5513..5842)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5527..5662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5581..5600)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5827..5847
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5856..6284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5966..5987)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6178..6197
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6330..6554
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6543..6560
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(6592..6978,6979..7593))
                     /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     7507..7526
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 cgaacaatct atccggcacg agttctctgt ggaatgcctt aacaacattg gccaggaaga
       61 aagggagcca gcacaatgta aagactccca tgataatgcc caaggtttta agcgctttct
      121 gttcccgcag ggcaacgagg cggcttggtc ttctttttcc agcccgacca ttggccagcg
      181 gtgcggtagc ggcagctgct gcgggtctag gtggtccggg tgggggggcg ggagcgggta
      241 cggggctggg tgacggggat gggggtctgg cgggaccacc cagaaacctc cgttcacaag
      301 aatcaatctt tttgacctgt ttctgtgcct cccggaaaac gcggagataa acgaaggcca
      361 tgatgcacaa aggcacgtag aaagatacga cactggacgc gatcgcgtaa gcgcgattgg
      421 tgacgaagtc acagcactta gggtcattgt aacaccttct agcctcatca gactctgctc
      481 gccaccaatg catgaggatc gggaggaagc tcaccagggc gctgatggcc cacacggtac
      541 agaccagtcc tctggcccgg gctcttgtca gcaggctttg atatctgaac ggactggtaa
      601 ttgcaaggta gcgatccagg gcgattacgc acagagtctc tattgatgcg gtaacgcaca
      661 acacatctac actagtccag agctcgcaga agaaggatcc atactcccat cgcccccaaa
      721 ccacgattgt agcgccgaat ggaaccacga gcagtcccat caccaggtcg gcggaagcca
      781 ggctcattat gaaaagattt gtgagggttt gaaggcgggg ggttttggcg atagcaacta
      841 tcaccaacac atttcctgct actataagca gcacaatcaa ggccatgagc agtcccatgc
      901 cagcggtcca ttgctggctc aggggctctg ggctctcact ggcgggcggg agcaatgacg
      961 caggggggct agccggaacc agcagtcttg cggctgtagc agctccatca ggcagaggtg
     1021 cagcagagct aagatttccc ggctcgctgg cccccagcac aagtactcct gcccccatat
     1081 cgatgctggc gtcatcatcg tccttgtagt cggcgaatac caggcagaag atgtagctca
     1141 gggcgatgat cgtcttcatg gtggcgcggc cgccactgtg ctggatatct gcagaattcc
     1201 accacactgg actagtggat ccgagctcgg taccaagctt aagtttaaac gctagccagc
     1261 ttgggtctcc ctatagtgag tcgtattaat ttcgataagc cagtaagcag tgggttctct
     1321 agttagccag agagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     1381 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     1441 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     1501 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     1561 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     1621 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     1681 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     1741 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     1801 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     1861 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacg
     1921 cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt
     1981 catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg cgcactactc
     2041 agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg catcagagca
     2101 gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc gacgtcaggt
     2161 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2221 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2281 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2341 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2401 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2461 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2521 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2581 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2641 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2701 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2761 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2821 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2881 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2941 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3121 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3181 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3961 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4021 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4081 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4141 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4201 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4261 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     4321 aagctctagc tagaggtcga cggtatacag acatgataag atacattgat gagtttggac
     4381 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4441 ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa gaactccagc
     4501 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
     4561 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
     4621 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
     4681 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     4741 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     4801 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     4861 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg
     4921 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     4981 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     5041 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     5101 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     5161 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     5221 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg
     5281 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     5341 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     5401 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     5461 atcctgtctc ttgatcagat ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa
     5521 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gcagaggcgg
     5581 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     5641 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     5701 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     5761 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5821 ttccacaccc taactgacac acattccaca gaattaattc gcgttaaatt tttgttaaat
     5881 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     5941 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6001 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6061 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     6121 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     6181 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     6241 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga
     6301 gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct
     6361 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     6421 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     6481 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     6541 actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag actacccacc
     6601 gtactcgtca attccaaggg catcggtaaa catctgctca aactcgaagt cggccatatc
     6661 cagagcgccg tagggggcgg agtcgtgggg ggtaaatccc ggacccgggg aatccccgtc
     6721 ccccaacatg tccagatcga aatcgtctag cgcgtcggca tgcgccatcg ccacgtcctc
     6781 gccgtctaag tggagctcgt cccccaggct gacatcggtc gggggggccg tcgacagtct
     6841 gcgcgtgtgt cccgcgggga gaaaggacag gcgcggagcc gccagccccg cctcttcggg
     6901 ggcgtcgtcg tccgggagat cgagcaggcc ctcgatggta gacccgtaat tgtttttcgt
     6961 acgcgcgcgg ctgtacgcgg acccactttc acatttaagt tgtttttcta atccgcatat
     7021 gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgatcaa ataattcgat
     7081 agcttgtcgt aataatggcg gcatactatc agtagtaggt gtttcccttt cttctttagc
     7141 gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca cagcgctgag
     7201 tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta attgattttc
     7261 gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg
     7321 atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat cttgccagct
     7381 ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa tggctaaggc
     7441 gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct ctacacctag
     7501 cttctgggcg agtttacggg ttgttaaacc ttcgattccg acctcattaa gcagctctaa
     7561 tgcgctgtta atcactttac ttttatctaa tcttagctgg aagtacaggt tctcaccggt
     7621 cgatgaagtg tccttggcca gggaggagct ggcggtggtg caggactcat cttggggacc
     7681 caggctgggt ggggtgcgtc caccggtatc gatgaccttg gactctgaag caaagccggg
     7741 tctacaaggt tcgtccaagg agctgtcaga atctgcagca gcgccgccat tgcaacctgc
     7801 ccagggctcc agcaggcgcg ctggtggtgt agctccgacc acatcgtcgt cgtcatcgtc
     7861 ggaggcagct cctggggagg gtggtgggcc cgggcgtgcg agacatccgg acgcccttgg
     7921 ccggtccccg tgagttgcgt gcctccttct agcggcccgg cgggcgcaac aaaggaggcc
     7981 ttggaaggcc tttctgaaat cgggagagcg gcagtagata atgggattga aagcagagtt
     8041 ggcgtatccc agccagttaa agaaga
//
