LOCUS       sequence-456479-        8255 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 8255)
  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 8255)
  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: GPR75-Tango
FEATURES             Location/Qualifiers
     source          1..8255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(166..234,235..1026)
                     /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     complement(384..403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1197..1785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1686..1705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1939..1956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2073..2094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2109..2114,2115..2132,2133..2139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2147..2163
                     /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)."
     primer_bind     2152..2174
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2171..2187
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2171..2187
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2236..2357
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2247..2266)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2301..2320
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2531..3325)
                     /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(2642..2661)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3252..3271
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3392..3721)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3406..3541
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3460..3479)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3706..3726
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3735..4163)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3845..3866)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4057..4076
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4209..4433
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4422..4439
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(4471..4857,4858..5472))
                     /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     5386..5405
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7403..7422)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7404..7422)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7467..7670)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7496..7516)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7671..8050
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     8222..8241
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taggggatcg ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg
       61 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      181 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      241 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      301 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      361 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      421 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      481 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      541 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      601 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      661 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      721 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      781 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      841 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      901 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      961 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1021 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1081 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1321 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1801 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1861 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1921 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1981 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2041 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     2101 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2161 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     2221 ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2281 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2341 gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     2401 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     2461 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     2521 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     2581 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     2641 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     2701 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     2761 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     2821 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     2881 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     2941 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3001 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3061 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3121 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     3181 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3241 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3301 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc
     3361 cgaaaatgga tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc
     3421 ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa
     3481 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     3541 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     3601 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     3661 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     3721 cattccacag aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3781 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3841 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3901 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3961 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4021 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4081 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4141 taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc
     4201 ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     4261 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     4321 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     4381 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
     4441 tgatcagcgg gtttaaacgg gccctctaga ctacccaccg tactcgtcaa ttccaagggc
     4501 atcggtaaac atctgctcaa actcgaagtc ggccatatcc agagcgccgt agggggcgga
     4561 gtcgtggggg gtaaatcccg gacccgggga atccccgtcc cccaacatgt ccagatcgaa
     4621 atcgtctagc gcgtcggcat gcgccatcgc cacgtcctcg ccgtctaagt ggagctcgtc
     4681 ccccaggctg acatcggtcg ggggggccgt cgacagtctg cgcgtgtgtc ccgcggggag
     4741 aaaggacagg cgcggagccg ccagccccgc ctcttcgggg gcgtcgtcgt ccgggagatc
     4801 gagcaggccc tcgatggtag acccgtaatt gtttttcgta cgcgcgcggc tgtacgcgga
     4861 cccactttca catttaagtt gtttttctaa tccgcatatg atcaattcaa ggccgaataa
     4921 gaaggctggc tctgcacctt ggtgatcaaa taattcgata gcttgtcgta ataatggcgg
     4981 catactatca gtagtaggtg tttccctttc ttctttagcg acttgatgct cttgatcttc
     5041 caatacgcaa cctaaagtaa aatgccccac agcgctgagt gcatataatg cattctctag
     5101 tgaaaaacct tgttggcata aaaaggctaa ttgattttcg agagtttcat actgtttttc
     5161 tgtaggccgt gtacctaaat gtacttttgc tccatcgcga tgacttagta aagcacatct
     5221 aaaactttta gcgttattac gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa
     5281 gtgagtatgg tgcctatcta acatctcaat ggctaaggcg tcgagcaaag cccgcttatt
     5341 ttttacatgc caatacaatg taggctgctc tacacctagc ttctgggcga gtttacgggt
     5401 tgttaaacct tcgattccga cctcattaag cagctctaat gcgctgttaa tcactttact
     5461 tttatctaat cttagctgga agtacaggtt ctcaccggtc gatgaagtgt ccttggccag
     5521 ggaggagctg gcggtggtgc aggactcatc ttggggaccc aggctgggtg gggtgcgtcc
     5581 accggtatcg atcacagaag gcaccggaat ctgcttggcg cttgtgctgt cgtactcttg
     5641 aaccagatca ttggttgtgt ggtagtgcat agcgatatag gagttggcga agccgaaact
     5701 gttcacaggc tggaggttac agggagaaga ctcctcctga gaaggggaac tgttgtatat
     5761 gctatagtat ggttctatgc gggtgttaat tggtgtggat gaagactgac cgcaatgttg
     5821 atggcctgct gaaatcttgg gagagaccat gctctccttg gaatggctag gaccgcaggc
     5881 ctgatccacg aacttcttct gtggcttggg tgacagcata tatgcgctgt ttgtttcgtg
     5941 gtgactactc ttattcctgt ttacttccag gttacccttg cccatcgcgc gcaatctggt
     6001 tttctgcttg caacagaaaa agccaaggcc gatatattgc aaacaccaca gtaccttccg
     6061 tctcaatcca gcagagtttc tgctgtagat aaatggattg aggccgctct tgaagaagat
     6121 cagtgtgaat ccaaacaatt cgaattggta caagataaat gatccatttg aagacagaac
     6181 cacttgcaca agtgaaatcc cgagcggcag gcaacacacc aggacagaga gtacaataat
     6241 gacacaagta accacggcct tactatcctt ggctgtagac aagttaatgg cggagaccag
     6301 ctgaagcctg gatgccgcgg gagtaacgag ctggttggga gacttcgtgt atcctctcgt
     6361 ctgcacatgc tgcagtttgt tatagttctg gttgcgatac agtgcaggca ttgcacactg
     6421 aattgggtct ccgcctccct gcaccggaac tcccatgaac ggctgaggcc gagaggcgtc
     6481 gactgtgatg acaggagggc attttctcac ctgagcgttt ttgcgcaaag tctgggctat
     6541 catgatgtaa gacactgaga cgacggccac gcaaaaggta aaatctacaa cgtaaagaga
     6601 cagaatagct ttacccttcc cggcgataag tgagctcatt ggaagacaca ggtgagactt
     6661 ggaagtcttc agggtcgcca gtgtggccag tgtaaagcta gtagcccaaa gaagcagagt
     6721 cagcagaacc gtacatggaa agctagcggt gcgattaggc tgcttgccaa gaaccatcct
     6781 gagtcggtgg agtgctatga ccgcgactgt cttcaaggac ataattatga agccgctgga
     6841 agtcagatgg aaagtgaagc aaaacgcatc gggaatactg gacgcgctac taaaaaaaag
     6901 aacgaaagta aacataggtg cggttactcc gcagatgaac aaatcgcaaa atgaaaggtt
     6961 caggatcata aaatcgaaat tggtccggaa cttcctgaag gcggggtcga aaaaagagag
     7021 aaaaacaata aagttcccat atgacccgag acagaagatc acagccaaca gaaacgtgca
     7081 ggttaccaaa gtggcggtgt gaataaggtc ctgcaagccc tcctgcaggc ttgtagaatt
     7141 tccttcctgt gagtgtggca catggagact tgtggcgtta ggagcgtcct gcagatgacc
     7201 agttgaattc atatcgatgc tggcgtcatc atcgtccttg tagtcggcga ataccaggca
     7261 gaagatgtag ctcagggcga tgatcgtctt catggtggcg cggccgccac tgtgctggat
     7321 atctgcagaa ttccaccaca ctggactagt ggatccgagc tcggtaccaa gcttaagttt
     7381 aaacgctagc cagcttgggt ctccctatag tgagtcgtat taatttcgat aagccagtaa
     7441 gcagtgggtt ctctagttag ccagagagct ctgcttatat agacctccca ccgtacacgc
     7501 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     7561 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     7621 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     7681 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     7741 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     7801 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     7861 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     7921 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     7981 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     8041 aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc
     8101 ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg
     8161 ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat
     8221 gcggcatcag agcagattgt actgagagtg cacca
//
