LOCUS       sequence-351271-        8474 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 8474)
  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 8474)
  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: GPR37-Tango
FEATURES             Location/Qualifiers
     source          1..8474
                     /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(2798..3412,3413..3799)
                     /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(2865..2884)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3831..3848)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3837..4061
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4107..4535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4194..4213)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4404..4425
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4544..4564)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4549..4878
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4729..4864
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4791..4810
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4945..5739
                     /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(4999..5018)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5609..5628
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5913..6034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5950..5969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6004..6023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6083..6099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6083..6099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6096..6118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6107..6123
                     /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(6131..6137,6138..6155,6156..6161))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6176..6197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6314..6331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6485..7073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6565..6584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7244..8035,8036..8104))
                     /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     7867..7886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8105..8209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8284..8303)
                     /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 gcgcgcccct ggcgctctgc tggcaaggat gagcagattg ttgctgctgc tcctgctgaa
      901 agtgtctgct tcctctgccc tcggtgtggc tcctgcttca aggaacgaaa cctgtctggg
      961 ggaatcatgc gcgccgacag tgatccagcg gcgcggtcgc gatgcatggg ggcctgggaa
     1021 tagcgcacgg gacgtcctta gagctcgcgc tcccagggag gagcagggcg cagctttcct
     1081 cgccggaccc tcatgggacc ttccagccgc acccggacgc gacccggccg ccggccgggg
     1141 agctgaagca tcagccgctg gaccacccgg tcccccaact cgccctcccg gtccctggag
     1201 atggaaaggg gcccgaggcc aagaaccatc agagactctg ggcaggggga atccaaccgc
     1261 tttgcagctg tttctgcaga tcagcgagga agaggagaaa ggcccgcgcg gtgccgggat
     1321 cagtggcaga agccaggagc agagcgtcaa aacagtccct ggcgccagtg acctgttcta
     1381 ctggcctaga agggctggca aactccaggg aagccaccac aaaccgctct ccaagaccgc
     1441 aaatggcctg gctggccatg aagggtggac aatcgccttg cccggccggg cactggccca
     1501 gaatgggtct ctgggagaag gtatccacga gccagggggg ccccgaagag ggaatagcac
     1561 caatcggaga gtgagactca agaacccttt ttacccgctt acgcaggaat cttacggagc
     1621 ctatgccgtc atgtgtctga gcgtggtgat attcggaacc ggaatcatcg gcaacctggc
     1681 agttatgtgt attgtctgcc ataactacta catgcgaagt atctccaact ccctgctcgc
     1741 caatctggcg ttttgggatt tcctgatcat ctttttttgc ctgccactgg ttatcttcca
     1801 cgaactgacc aagaaatggt tgctggagga tttctcctgc aagattgtac cctacattga
     1861 agtggcttct cttggagtca ctactttcac tctgtgcgcc ctctgtatag atagatttag
     1921 ggccgccact aatgtacaga tgtattacga aatgatcgag aactgcagct ccacgacagc
     1981 caaactggcc gttatctggg tgggtgcatt gctcctggcg cttccagaag ttgtgctgcg
     2041 gcagctgagc aaggaggacc tcggcttcag tggcagagcg ccagctgaga gatgcatcat
     2101 caaaatcagc cccgacctgc ctgacacaat ttacgtgttg gcccttacct acgactctgc
     2161 gcggctttgg tggtacttcg gttgttactt ttgccttccc acactgttta ccatcacttg
     2221 ttcactggtc accgcccgga aaatcaggaa agctgagaaa gcctgcacta gaggcaacaa
     2281 gcgccaaatt cagcttgaat cacagatgaa ttgtacggtt gtggcactga ccattttgta
     2341 tggcttctgt atcatcccgg agaacatctg taacatcgtc actgcttaca tggcaaccgg
     2401 ggtctcccag cagaccatgg acttgctcaa tataatctcc caatttctct tgttctttaa
     2461 gagctgtgtc acccccgtcc tgctgttctg cctttgcaag cccttcagtc gcgcattcat
     2521 ggaatgttgc tgctgctgct gcgaggaatg cattcaaaaa tcctccacag taacatctga
     2581 cgacaatgat aatgagtaca ccaccgagct tgaactttct ccctttagca caatccggcg
     2641 ggaaatgtcc acctttgcat ctgtcggcac ccattgcatc gataccggtg gacgcacccc
     2701 acccagcctg ggtccccaag atgagtcctg caccaccgcc agctcctccc tggccaagga
     2761 cacttcatcg accggtgaga acctgtactt ccagctaaga ttagataaaa gtaaagtgat
     2821 taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact
     2881 cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc
     2941 tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt
     3001 agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt
     3061 actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta
     3121 tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc
     3181 attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga
     3241 gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt
     3301 acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct
     3361 tgaattgatc atatgcggat tagaaaaaca acttaaatgt gaaagtgggt ccgcgtacag
     3421 ccgcgcgcgt acgaaaaaca attacgggtc taccatcgag ggcctgctcg atctcccgga
     3481 cgacgacgcc cccgaagagg cggggctggc ggctccgcgc ctgtcctttc tccccgcggg
     3541 acacacgcgc agactgtcga cggccccccc gaccgatgtc agcctggggg acgagctcca
     3601 cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga
     3661 catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta
     3721 cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat
     3781 tgacgagtac ggtgggtagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3841 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3901 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3961 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4021 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4081 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4141 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4201 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4261 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4321 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4381 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4441 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4501 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4561 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4621 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4681 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4741 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4801 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4861 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4921 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4981 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5041 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5101 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5161 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5221 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5281 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5341 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5401 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5461 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5521 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5581 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5641 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5701 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5761 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5821 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5881 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5941 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6001 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6061 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6121 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6181 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6241 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8221 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     8281 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     8341 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     8401 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     8461 cagatatacg cgtt
//
