LOCUS       sequence-157894-        8192 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 8192)
  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 8192)
  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: ADRA1B-Tango
FEATURES             Location/Qualifiers
     source          1..8192
                     /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(2753..3367,3368..3754)
                     /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(2820..2839)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3786..3803)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3792..4016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4062..4490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4149..4168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4359..4380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4499..4519)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4504..4833
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4684..4819
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4746..4765
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4900..5694
                     /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(4954..4973)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5564..5583
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5868..5989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5905..5924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5959..5978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6038..6054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6038..6054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6051..6073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6062..6078
                     /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(6086..6092,6093..6110,6111..6116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6131..6152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6269..6286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6440..7025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6520..6539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7196..7987,7988..8056))
                     /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     7819..7838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8057..8161)
                     /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 atatgaaccc
     1081 agacttggac acaggccata acacaagtgc acccgcgcac tggggggagc tgaagaatgc
     1141 gaattttacc ggccctaatc agacaagctc caacagcact ctgccacagt tggatatcac
     1201 gcgcgctatc tcagttgggc tggttctcgg agccttcatc ctcttcgcca tcgtcggcaa
     1261 tatactggtg atcctctcag tggcatgtaa taggcatttg aggactccta ccaattattt
     1321 tattgtgaac ttggctatgg ctgacttgct tctgtcattc acagtgcttc cattcagtgc
     1381 agccttggag gtgctggggt actgggtgtt ggggagaatc ttttgtgaca tatgggcagc
     1441 cgtggacgtg ctgtgctgca ctgccagtat cttgtctttg tgcgcgatct ctatagatcg
     1501 gtacataggt gtcagatata gtcttcagta tccaacactc gtaactcgcc gcaaagcaat
     1561 tctcgccctg ctgagcgttt gggtcttgtc taccgttatc tccattggac cgctgttggg
     1621 atggaaggag ccagctccca acgatgacaa ggagtgtgga gtgacagaag aaccttttta
     1681 tgcactgttc agcagccttg gcagctttta tattccactt gccgtcatcc tggttatgta
     1741 ctgccgggtt tatattgtag ccaagcggac aaccaaaaac ctggaagccg gggtcatgaa
     1801 ggagatgtct aatagcaagg agctgacctt gcggatccac agcaaaaatt ttcatgagga
     1861 tactctttct agtactaagg caaaaggaca taaccctcgc agtagcattg ccgtgaagct
     1921 gttcaaattt agtcgagaga aaaaggcggc caaaaccctg ggcatcgtcg tgggcatgtt
     1981 tattctttgt tggttgccct tcttcatagc cctccctttg ggaagcctgt tctccaccct
     2041 gaaaccacca gacgccgtgt tcaaggtcgt cttttggttg ggctacttta acagttgcct
     2101 caatccaata atctatccct gctcatcaaa agaattcaag agggcctttg tgcgcatcct
     2161 gggctgtcaa tgccggggaa ggggtaggag acgacgcaga cggcggagac gccttggagg
     2221 gtgcgcgtat acctatagac cctggactag aggtggcagt ttggagagaa gccagtctcg
     2281 gaaagatagt ttggatgact ccggtagctg tttgagcgga tcccagagga ccctgccaag
     2341 cgcatctcca tcaccagggt acctgggccg gggtgccccc cctcccgttg aactgtgcgc
     2401 cttccctgaa tggaaagctc caggcgcact gctgagtctg ccagctcctg agcctccagg
     2461 ccggcggggg agacatgata gtgggccctt gttcactttc aaactgctta cagaaccgga
     2521 gtctcctgga acagatggcg gcgccagcaa cggaggttgc gaggccgccg ctgacgtggc
     2581 taacggccag cctggtttca aaagcaacat gccgttggcg ccaggccaat tcatcgatac
     2641 cggtggacgc accccaccca gcctgggtcc ccaagatgag tcctgcacca ccgccagctc
     2701 ctccctggcc aaggacactt catcgaccgg tgagaacctg tacttccagc taagattaga
     2761 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     2821 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
     2881 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     2941 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
     3001 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
     3061 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     3121 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
     3181 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     3241 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     3301 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     3361 tgggtccgcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca tcgagggcct
     3421 gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc cgcgcctgtc
     3481 ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg atgtcagcct
     3541 gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg acgcgctaga
     3601 cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat ttacccccca
     3661 cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc agatgtttac
     3721 cgatgccctt ggaattgacg agtacggtgg gtagtctaga gggcccgttt aaacccgctg
     3781 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3841 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3901 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3961 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4021 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4081 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4141 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4201 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4261 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4321 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4381 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4441 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4501 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4561 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4621 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4681 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4741 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4801 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4861 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4921 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4981 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5041 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5101 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5161 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5221 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5281 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5341 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5401 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5461 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5521 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5581 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5641 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5701 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5761 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5821 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5881 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5941 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6001 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6061 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6121 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6181 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6241 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6301 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6361 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6421 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6481 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6541 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6601 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6661 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6721 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6781 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6841 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6901 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6961 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7081 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7141 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7201 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7261 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7321 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7381 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7441 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7501 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7561 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7621 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7681 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7741 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7801 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7861 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7921 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7981 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8041 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8101 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8161 gcacatttcc ccgaaaagtg ccacctgacg tc
//
