LOCUS       sequence-358136-        7730 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 7730)
  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 7730)
  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: GPR68-Tango
FEATURES             Location/Qualifiers
     source          1..7730
                     /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(2288..2902,2903..3289)
                     /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(2355..2374)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3321..3338)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3327..3551
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3597..4025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3684..3703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3894..3915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4034..4054)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4039..4368
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4219..4354
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4281..4300
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4435..5229
                     /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(4489..4508)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5099..5118
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5403..5524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5440..5459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5494..5513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5573..5589)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5573..5589)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5586..5608)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5597..5613
                     /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(5621..5627,5628..5645,5646..5651))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5666..5687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5804..5821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5975..6563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6055..6074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6734..7525,7526..7594))
                     /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     7357..7376
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7595..7699)
                     /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 atatggggaa
     1081 catcactgcc gacaatagct ccatgagctg taccatcgac cacactatcc accagaccct
     1141 cgctcccgtg gtttacgtta cggtacttgt cgtaggcttc cctgccaact gcctcagtct
     1201 ctactttggg tacctgcaga taaaagctcg caacgagttg ggagtgtact tgtgcaacct
     1261 gacagtggca gacctgttct acatatgctc ccttcccttt tggctgcagt acgtcctcca
     1321 gcatgataat tggtcccatg gtgacctgag ctgccaagtg tgcgggattc ttctttatga
     1381 gaatatctac atttccgtgg gattcctttg ctgcatcagc gtggaccgct acctggccgt
     1441 agcccatcct ttccgattcc accagttccg gacactcaag gctgcagtgg gggttagcgt
     1501 ggtgatctgg gctaaagaac tcctgacctc aatatacttc ttgatgcacg aggaagtgat
     1561 cgaggacgag aatcagcaca gggtctgctt cgagcactac cctattcaag cttggcagcg
     1621 agctatcaat tactacagat ttctcgtggg gttcctgttt cctatctgtc tgctgctggc
     1681 ctcctaccag ggaatcctgc gagccgttcg ccgcagccat ggtactcaga aatctcgcaa
     1741 agaccaaatt cagagacttg tgctcagcac tgtggtcatt ttcctcgcgt gcttcctgcc
     1801 atatcatgtg ttgctgctgg tcagaagcgt gtgggaggca tcttgtgact tcgccaaggg
     1861 agtctttaac gcctatcatt tcagtctcct tctgacatct ttcaattgtg tcgccgatcc
     1921 tgtgctctat tgcttcgttt ccgaaacaac tcatagggat cttgcacgcc tccgcggagc
     1981 ttgcctggcc ttccttacat gttcaaggac cggcagggca cgcgaagcat atccgctggg
     2041 tgccccagag gcttctggta agtcaggcgc gcaaggagag gagccggaac tgcttaccaa
     2101 gctccacccc gcttttcaaa cccccaactc ccccgggtcc ggcggctttc caaccgggag
     2161 acttgcaatc gataccggtg gacgcacccc acccagcctg ggtccccaag atgagtcctg
     2221 caccaccgcc agctcctccc tggccaagga cacttcatcg accggtgaga acctgtactt
     2281 ccagctaaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt
     2341 cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac
     2401 attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt
     2461 agataggcac catactcact tttgcccttt agaaggggaa agctggcaag attttttacg
     2521 taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca
     2581 tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt
     2641 atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg tggggcattt
     2701 tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac
     2761 acctactact gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca
     2821 aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca
     2881 acttaaatgt gaaagtgggt ccgcgtacag ccgcgcgcgt acgaaaaaca attacgggtc
     2941 taccatcgag ggcctgctcg atctcccgga cgacgacgcc cccgaagagg cggggctggc
     3001 ggctccgcgc ctgtcctttc tccccgcggg acacacgcgc agactgtcga cggccccccc
     3061 gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca
     3121 tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc
     3181 gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg acttcgagtt
     3241 tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtgggtagt ctagagggcc
     3301 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3361 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3421 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3481 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3541 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3601 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3661 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3721 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3781 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3841 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3901 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3961 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4021 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4141 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4201 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4261 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4321 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4381 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4441 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4501 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4561 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4621 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4681 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4741 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4801 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4861 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4921 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4981 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5041 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5101 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5161 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5221 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5281 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5341 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5401 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5461 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5521 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5581 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5641 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5701 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5761 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5821 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5881 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5941 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6001 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6061 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6121 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6181 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6241 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6301 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6361 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6421 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6481 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6541 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6601 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6661 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6721 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6781 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6841 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6901 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6961 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7021 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7081 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7141 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7201 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7261 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7321 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7381 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7441 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7501 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7561 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7621 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7681 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
