LOCUS       sequence-344664-        7889 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 7889)
  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 7889)
  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: NTSR1-Tango
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2213..2827,2828..3214)
                     /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(2280..2299)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3246..3263)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3252..3476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3522..3950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3609..3628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3819..3840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3959..3979)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3964..4293
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4144..4279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4206..4225
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4360..5154
                     /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(4414..4433)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5024..5043
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5328..5449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5365..5384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5419..5438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5498..5514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5498..5514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5511..5533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5522..5538
                     /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(5546..5552,5553..5570,5571..5576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5591..5612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5729..5746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5900..6488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5980..5999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6659..7450,7451..7519))
                     /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     7282..7301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7520..7624)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7699..7718)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttctgcagat atccagcaca gtggcggccg cgccaccatg aagacgatca tcgccctgag
      781 ctacatcttc tgcctggtat tcgccgacta caaggacgat gatgacgcca gcatcgatat
      841 gagactgaac tcttccgctc caggaacgcc gggcacaccg gctgcagatc ccttccaaag
      901 ggcccaggcc ggcttggaag aggccctgct cgctccgggc ttcggaaacg cctctggaaa
      961 tgcctctgaa cgcgtgcttg ccgcaccctc cagtgagctc gatgttaaca ccgacatcta
     1021 cagcaaagtc ctggtgaccg cggtatatct ggcactcttc gtcgttggca ctgtaggtaa
     1081 cacggtgact gcttttaccc ttgcgcggaa gaagagcctt cagagcctgc aaagcacagt
     1141 acattaccat ctgggttccc tggccctttc tgacctgctc acactcctgc tggctatgcc
     1201 agtggaactg tacaacttca tctgggtgca ccatccttgg gcatttgggg acgctggctg
     1261 tcgcgggtat tattttctta gagacgcctg cacttatgcc accgccttga atgtagcctc
     1321 tctcagcgta gagagatacc tcgctatctg ccatcccttc aaggctaaga ccttgatgag
     1381 ccgaagccga actaaaaagt tcatctcagc aatttggctt gcgagcgcac tgctggccgt
     1441 tcctatgctg tttactatgg gtgagcagaa tagaagcgca gatggacaac acgccggagg
     1501 cctggtgtgc acacccacca ttcacactgc cacggtgaaa gtggtcattc aggtgaacac
     1561 attcatgagc tttatctttc ccatggtcgt aatcagcgtt ctcaacacca taattgcaaa
     1621 taagctgaca gtgatggtgc ggcaggccgc tgagcaagga caggtatgta ccgttggcgg
     1681 cgagcactca acattcagca tggcaattga acccgggaga gtccaggcac tgagacatgg
     1741 agtgcgggtt ctgcgggcgg tggtgatcgc gttcgtagtt tgttggctcc cgtaccacgt
     1801 ccggcgcctc atgttctgct atatttccga cgaacagtgg acacctttcc tttacgactt
     1861 ttaccattac ttctatatgg tgacgaacgc tcttttctat gtttccagta caatcaatcc
     1921 tatcctttac aacctggtca gtgcaaactt ccgacatatc ttcctggcaa cactcgcctg
     1981 tctgtgcccc gtttggcgcc gccggaggaa gcgccccgcc ttctcacgga aggccgactc
     2041 agtatcctct aaccataccc tgagttcaaa cgccactcgg gaaacgctgt acatcgatac
     2101 cggtggacgc accccaccca gcctgggtcc ccaagatgag tcctgcacca ccgccagctc
     2161 ctccctggcc aaggacactt catcgaccgg tgagaacctg tacttccagc taagattaga
     2221 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     2281 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
     2341 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     2401 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
     2461 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
     2521 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     2581 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
     2641 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     2701 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     2761 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     2821 tgggtccgcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca tcgagggcct
     2881 gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc cgcgcctgtc
     2941 ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg atgtcagcct
     3001 gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg acgcgctaga
     3061 cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat ttacccccca
     3121 cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc agatgtttac
     3181 cgatgccctt ggaattgacg agtacggtgg gtagtctaga gggcccgttt aaacccgctg
     3241 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3301 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3361 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3421 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3481 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3541 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3601 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3661 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3721 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3781 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3841 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3901 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3961 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4021 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4141 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4201 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4261 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4321 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4381 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4441 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4501 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4561 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4621 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4681 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4741 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4801 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4861 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4921 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4981 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5041 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5101 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5161 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5221 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5281 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5341 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5401 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5461 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5521 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5581 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5641 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5701 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6301 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6601 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6661 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6721 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6781 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6841 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6901 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6961 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7021 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7081 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7141 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7201 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7261 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7321 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7381 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7441 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7501 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7561 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7621 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7681 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7741 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     7801 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     7861 tcgcgatgta cgggccagat atacgcgtt
//
