LOCUS       sequence-430904-        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The DuET construct encodes the named human GPCR engineered with a 
            cleaved hemagglutinin signal peptide, a 5' FLAG and a 3' 1D4 epitope
            tag optimized for expression in mammalian cells and proteomics 
            studies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  AUTHORS   Dahl L, Kotliar IB, Bendes A, Dodig-Crnkovic T, Fromm S, Elofsson A,
            Uhlen M, Sakmar TP, Schwenk JM
  TITLE     Multiplexed selectivity screening of anti-GPCR antibodies.
  JOURNAL   Sci Adv. 2023 May 3;9(18):eadf9297. doi: 10.1126/sciadv.adf9297. 
            Epub 2023 May 3.
  PUBMED    37134173
REFERENCE   2  (bases 1 to 7301)
  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"; journalName: "Sci Adv."; 
            date: "2023-05-3"; volume: "9(18):eadf9297. doi"; pages: " 
            10.1126/sciadv.adf9297. Epub 2023 May 3"
COMMENT     Sequence Label: GPR37-DuET
FEATURES             Location/Qualifiers
     source          1..7301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(133..237)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(312..331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        503..882
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        883..1086
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1037..1057
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1131..1150
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1131..1149
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3160..3177)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3166..3390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3436..3864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3523..3542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3733..3754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3873..3893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3878..4207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4120..4139
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4274..5068
                     /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(4328..4347)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4938..4957
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5242..5363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5279..5298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5333..5352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5412..5428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5412..5428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5425..5447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5436..5452
                     /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(5460..5466,5467..5484,5485..5490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5505..5526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5643..5660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5814..6402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5894..5913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6573..7301,1..63,64..132))
                     /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     7196..7215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
       61 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      121 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      181 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      241 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      301 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      361 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      421 ttgaccgaca tttgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      481 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      541 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      601 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      661 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      721 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      781 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      841 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      901 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      961 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1021 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1081 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     1141 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttagcggc cgcgccacca
     1201 tgaagacgat catcgccctg agctacatct tctgcctggt attcgccgac tacaaggacg
     1261 atgatgacgc cagcatcgat gccctcggtg tggctcctgc ttcaaggaac gaaacctgtc
     1321 tgggggaatc atgcgcgccg acagtgatcc agcggcgcgg tcgcgatgca tgggggcctg
     1381 ggaatagcgc acgggacgtc cttagagctc gcgctcccag ggaggagcag ggcgcagctt
     1441 tcctcgccgg accctcatgg gaccttccag ccgcacccgg acgcgacccg gccgccggcc
     1501 ggggagctga agcatcagcc gctggaccac ccggtccccc aactcgccct cccggtccct
     1561 ggagatggaa aggggcccga ggccaagaac catcagagac tctgggcagg gggaatccaa
     1621 ccgctttgca gctgtttctg cagatcagcg aggaagagga gaaaggcccg cgcggtgccg
     1681 ggatcagtgg cagaagccag gagcagagcg tcaaaacagt ccctggcgcc agtgacctgt
     1741 tctactggcc tagaagggct ggcaaactcc agggaagcca ccacaaaccg ctctccaaga
     1801 ccgcaaatgg cctggctggc catgaagggt ggacaatcgc cttgcccggc cgggcactgg
     1861 cccagaatgg gtctctggga gaaggtatcc acgagccagg ggggccccga agagggaata
     1921 gcaccaatcg gagagtgaga ctcaagaacc ctttttaccc gcttacgcag gaatcttacg
     1981 gagcctatgc cgtcatgtgt ctgagcgtgg tgatattcgg aaccggaatc atcggcaacc
     2041 tggcagttat gtgtattgtc tgccataact actacatgcg aagtatctcc aactccctgc
     2101 tcgccaatct ggcgttttgg gatttcctga tcatcttttt ttgcctgcca ctggttatct
     2161 tccacgaact gaccaagaaa tggttgctgg aggatttctc ctgcaagatt gtaccctaca
     2221 ttgaagtggc ttctcttgga gtcactactt tcactctgtg cgccctctgt atagatagat
     2281 ttagggccgc cactaatgta cagatgtatt acgaaatgat cgagaactgc agctccacga
     2341 cagccaaact ggccgttatc tgggtgggtg cattgctcct ggcgcttcca gaagttgtgc
     2401 tgcggcagct gagcaaggag gacctcggct tcagtggcag agcgccagct gagagatgca
     2461 tcatcaaaat cagccccgac ctgcctgaca caatttacgt gttggccctt acctacgact
     2521 ctgcgcggct ttggtggtac ttcggttgtt acttttgcct tcccacactg tttaccatca
     2581 cttgttcact ggtcaccgcc cggaaaatca ggaaagctga gaaagcctgc actagaggca
     2641 acaagcgcca aattcagctt gaatcacaga tgaattgtac ggttgtggca ctgaccattt
     2701 tgtatggctt ctgtatcatc ccggagaaca tctgtaacat cgtcactgct tacatggcaa
     2761 ccggggtctc ccagcagacc atggacttgc tcaatataat ctcccaattt ctcttgttct
     2821 ttaagagctg tgtcaccccc gtcctgctgt tctgcctttg caagcccttc agtcgcgcat
     2881 tcatggaatg ttgctgctgc tgctgcgagg aatgcattca aaaatcctcc acagtaacat
     2941 ctgacgacaa tgataatgag tacaccaccg agcttgaact ttctcccttt agcacaatcc
     3001 ggcgggaaat gtccaccttt gcatctgtcg gcacccattg catcgatacc ggtggacgca
     3061 ccggtgaatt cgatgaggcc agcaccaccg tgtctaagac cgagacttct caggtggccc
     3121 ctgcctaatc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3181 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3241 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3301 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3361 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3421 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3481 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3541 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3601 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3661 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3721 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3781 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3841 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4201 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4261 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4321 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4381 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4441 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4501 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4561 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4621 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4681 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4741 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4801 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4861 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4921 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4981 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5041 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5101 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5161 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5221 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5341 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5401 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5461 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5521 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5581 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5641 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5701 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5761 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5821 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5881 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5941 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6001 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6061 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6121 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6181 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6241 ggctacacta gaagaacggt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6301 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6361 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6421 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6481 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6541 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6601 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6661 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6721 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6781 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6841 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6901 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6961 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7021 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7081 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7141 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7201 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7261 aaactctcaa ggatcttacc gctgttgaga tccagttcga t
//
