LOCUS       sequence-456518-        7760 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 7760)
  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 7760)
  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: GPR27-Tango
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(664..1455,1456..1524))
                     /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     1287..1306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1525..1629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1704..1723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1895..2274
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2275..2478
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2429..2449
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2523..2542
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2523..2541
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3978..4592,4593..4979)
                     /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(4045..4064)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(5011..5028)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5017..5241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5287..5715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5374..5393)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5584..5605
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5724..5744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5729..6058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5909..6044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5971..5990
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6125..6919
                     /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(6179..6198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6789..6808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7093..7214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7130..7149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7184..7203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7263..7279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7263..7279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7276..7298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7287..7303
                     /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(7311..7317,7318..7335,7336..7341))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7356..7377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7494..7511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7665..7760,1..493))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(join(7745..7760,1..4))
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
       61 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
      541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
      601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
      781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
      841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
      901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
      961 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1021 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1261 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1621 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     1681 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     1741 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     1801 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     1861 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     1921 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1981 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2041 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2101 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2161 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2221 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2281 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     2341 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2401 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2461 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     2521 attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttggta
     2581 ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg
     2641 gccgcgccac catgaagacg atcatcgccc tgagctacat cttctgcctg gtattcgccg
     2701 actacaagga cgatgatgac gccagcatcg atatggccaa tgccagtgag ccaggcggct
     2761 ccgggggtgg cgaggccgca gccctggggt tgaaactggc cacactgtcc ctgcttcttt
     2821 gcgtgagcct cgctggtaac gtgctgttcg cactgctcat cgtgcgcgag aggtcactgc
     2881 accgcgcacc ctattacctg cttctcgatc tgtgtctggc cgacggtctc agggctttgg
     2941 catgcttgcc ggccgtcatg ctcgccgcga gaagggctgc tgcagcagcc ggagcaccac
     3001 ccggagcact cggctgcaaa ctgttggctt tcctcgccgc cctcttctgt tttcatgctg
     3061 cttttctcct cctgggcgtg ggcgtcacta gatatctggc tatcgcccac caccgattct
     3121 acgccgaacg gctggcgggt tggccatgtg ctgcaatgct tgtctgtgcc gcctgggcac
     3181 ttgctctggc tgcagcattt ccccctgtgc tggacggagg gggggacgat gaggatgcgc
     3241 catgcgcact ggaacagcgg cccgacggag ctcctggcgc cctcgggttt ctgctcttgc
     3301 tggctgtagt ggtcggggcc acacacttgg tatatttgcg gctgctgttc ttcatccacg
     3361 acagacgcaa gatgcggcct gccaggctgg tgccagcggt gtcccacgat tggacgttcc
     3421 acgggcctgg cgcaacgggt caggctgcag ccaactggac agccggattt ggaagaggac
     3481 ctaccccccc cgctctggtg ggaattaggc ccgccggacc aggaagggga gcccgcaggc
     3541 ttctggttct cgaagaattt aagactgaaa agcgactgtg caaaatgttt tacgcggtga
     3601 cgctgttgtt cctcctgctg tggggtccat atgtggtggc ctcttatctc agagtgctgg
     3661 tgaggccagg agctgtacct caggcctacc tcaccgccag cgtatggctg actttcgcgc
     3721 aggccggaat taatcctgtg gtgtgctttc tttttaacag ggaactccgg gattgcttcc
     3781 gggcacaatt cccatgctgc cagtccccga ggaccacaca ggcaactcac ccatgcgact
     3841 tgaaaggcat cggtctgatc gataccggtg gacgcacccc acccagcctg ggtccccaag
     3901 atgagtcctg caccaccgcc agctcctccc tggccaagga cacttcatcg accggtgaga
     3961 acctgtactt ccagctaaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     4021 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     4081 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     4141 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     4201 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     4261 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     4321 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
     4381 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     4441 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     4501 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     4561 tagaaaaaca acttaaatgt gaaagtgggt ccgcgtacag ccgcgcgcgt acgaaaaaca
     4621 attacgggtc taccatcgag ggcctgctcg atctcccgga cgacgacgcc cccgaagagg
     4681 cggggctggc ggctccgcgc ctgtcctttc tccccgcggg acacacgcgc agactgtcga
     4741 cggccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg
     4801 cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt
     4861 ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg
     4921 acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtgggtagt
     4981 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     5041 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5101 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5161 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     5221 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     5281 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5341 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5401 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5461 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5521 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5581 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5641 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5701 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5761 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5821 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5881 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5941 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     6001 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     6061 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     6121 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     6181 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     6241 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     6301 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     6361 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     6421 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     6481 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     6541 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6601 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     6661 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6721 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6781 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6841 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6901 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6961 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     7021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     7081 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7201 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     7261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7321 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7741 ctataaagat accaggcgtt
//
