LOCUS       sequence-456467-        7805 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 7805)
  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 7805)
  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: PTGER3-Tango
FEATURES             Location/Qualifiers
     source          1..7805
                     /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(2363..2977,2978..3364)
                     /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(2430..2449)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3396..3413)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3402..3626
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3672..4100
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3759..3778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3969..3990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4109..4129)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4114..4443
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4294..4429
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4356..4375
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4510..5304
                     /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(4564..4583)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5174..5193
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5478..5599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5515..5534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5569..5588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5648..5664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5648..5664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5661..5683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5672..5688
                     /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(5696..5702,5703..5720,5721..5726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5741..5762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5879..5896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6050..6638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6130..6149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6809..7600,7601..7669))
                     /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     7432..7451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7670..7774)
                     /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 atatgaaaga
     1081 gacgcgcggc tacggcggcg atgccccttt ctgtactaga ctcaatcaca gctacactgg
     1141 gatgtgggcc cctgaacgct ccgcagaggc tcgcgggaac ctgactcgcc cgccaggaag
     1201 tggggaagac tgcgggtctg tcagcgtggc atttcctata acaatgctgc tcactggctt
     1261 tgttgggaat gctctcgcca tgcttctcgt gagtaggagc taccgccgaa gggagtccaa
     1321 gcgcaagaaa agcttcctcc tttgtatcgg gtggctcgcc ctcacagact tggtaggtca
     1381 gcttctgact acacctgtag tcattgtggt gtatctctca aagcagagat gggaacatat
     1441 cgacccatcc ggccgacttt gtactttctt cggacttact atgactgtgt tcgggctgag
     1501 ctcactgttc attgctagcg ctatggccgt agagcgggcc ttggctatca gagcacctca
     1561 ttggtacgcc agtcatatga aaacccgcgc cacacgagct gtcctcctcg gcgtatggct
     1621 ggccgtgttg gcgttcgcac ttctgcctgt gctgggcgtg ggacagtaca cagtgcaatg
     1681 gccaggaaca tggtgcttca ttagcacagg gagaggtggg aacggaacaa gttcctctca
     1741 caactggggg aatctgttct ttgcctcagc attcgctttc ctcgggctgt tggccctcac
     1801 cgtcacattc tcatgtaacc tcgcaacaat caaggcactc gtgtctcgat gtagagcaaa
     1861 ggcaacagcc tcccagtcca gtgctcaatg gggccgcatc acaactgaaa ctgccatcca
     1921 gctgatgggg atcatgtgcg tgttgagcgt atgctggtca ccactgctga ttatgatgct
     1981 gaagatgatc ttcaaccaaa ccagcgtcga gcactgcaag actcataccg agaaacagaa
     2041 agaatgtaac tttttcctta tcgcggtgcg gctggctagt ctgaatcaga ttctcgaccc
     2101 ttgggtgtat ctcctgcttc gaaaaatttt gctgaggaag ttttgtcaaa tccgctacca
     2161 cacaaataat tatgcatcca gtagcacttc cctcccgtgt cagtgtagca gcaccctgat
     2221 gtggtccgac catctcgaac ggatcgatac cggtggacgc accccaccca gcctgggtcc
     2281 ccaagatgag tcctgcacca ccgccagctc ctccctggcc aaggacactt catcgaccgg
     2341 tgagaacctg tacttccagc taagattaga taaaagtaaa gtgattaaca gcgcattaga
     2401 gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc agaagctagg
     2461 tgtagagcag cctacattgt attggcatgt aaaaaataag cgggctttgc tcgacgcctt
     2521 agccattgag atgttagata ggcaccatac tcacttttgc cctttagaag gggaaagctg
     2581 gcaagatttt ttacgtaata acgctaaaag ttttagatgt gctttactaa gtcatcgcga
     2641 tggagcaaaa gtacatttag gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa
     2701 tcaattagcc tttttatgcc aacaaggttt ttcactagag aatgcattat atgcactcag
     2761 cgctgtgggg cattttactt taggttgcgt attggaagat caagagcatc aagtcgctaa
     2821 agaagaaagg gaaacaccta ctactgatag tatgccgcca ttattacgac aagctatcga
     2881 attatttgat caccaaggtg cagagccagc cttcttattc ggccttgaat tgatcatatg
     2941 cggattagaa aaacaactta aatgtgaaag tgggtccgcg tacagccgcg cgcgtacgaa
     3001 aaacaattac gggtctacca tcgagggcct gctcgatctc ccggacgacg acgcccccga
     3061 agaggcgggg ctggcggctc cgcgcctgtc ctttctcccc gcgggacaca cgcgcagact
     3121 gtcgacggcc cccccgaccg atgtcagcct gggggacgag ctccacttag acggcgagga
     3181 cgtggcgatg gcgcatgccg acgcgctaga cgatttcgat ctggacatgt tgggggacgg
     3241 ggattccccg ggtccgggat ttacccccca cgactccgcc ccctacggcg ctctggatat
     3301 ggccgacttc gagtttgagc agatgtttac cgatgccctt ggaattgacg agtacggtgg
     3361 gtagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3421 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3481 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3541 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3601 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3661 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3721 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3781 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3841 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3901 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3961 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4021 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4081 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4321 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4381 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4441 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4501 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4561 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4621 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4681 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4741 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4801 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4861 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4921 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4981 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5041 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5101 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5161 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5221 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5281 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5341 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5401 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5461 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5521 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5581 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5641 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5701 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5761 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5821 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5881 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5941 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6001 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6061 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6121 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6241 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6421 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6481 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7801 acgtc
//
