LOCUS       sequence-431807-        7988 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 7988)
  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 7988)
  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: GPR156-DuET
FEATURES             Location/Qualifiers
     source          1..7988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(122..226)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(301..320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        492..871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        872..1075
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1026..1046
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1120..1139
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1120..1138
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3836..3853)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3842..4066
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4112..4540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4199..4218)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4409..4430
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4549..4569)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4554..4883
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4734..4869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4796..4815
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4950..5744
                     /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(5004..5023)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5614..5633
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5918..6039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5955..5974)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6009..6028
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6088..6104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6088..6104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6101..6123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6112..6128
                     /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(6136..6142,6143..6160,6161..6166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6181..6202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6319..6336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6490..7078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6570..6589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7249..7988,1..52,53..121))
                     /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     7872..7891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
       61 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
      121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
      181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      241 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
      301 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
      361 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacat
      421 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
      481 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      541 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      601 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      661 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      721 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      781 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      841 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      901 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      961 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     1021 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     1081 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     1141 gacccaagct ggctagcgtt taaacttaag cttagcggcc gcgccaccat gaagacgatc
     1201 atcgccctga gctacatctt ctgcctggta ttcgccgact acaaggacga tgatgacgcc
     1261 agcatcgata tggagcccga gatcaattgc tccgagctgt gcgactcttt ccccggccag
     1321 gagctcgata gaagaccact gcatgacctc tgcaaaacta cgatcaccag cagtcaccat
     1381 tcaagtaaga ctatttcctc cctcagccct gtcctgctgg gcattgtttg gacgttcctc
     1441 tcatgcggtc tcttgttgat tttgtttttc ctggcattca ccatccactg cagaaagaat
     1501 agaatcgtga aaatgagcag ccctaacctt aacatcgtga cccttctggg aagctgcctg
     1561 acttatagta gcgcctacct ttttggcatc caggatgttc ttgtgggaag ctctatggag
     1621 accctgatac agactcgcct cagcatgctg tgtattggta cctccttggt cttcggacct
     1681 atccttggga agagctggag actgtacaag gttttcacgc agagagtgcc agataaaagg
     1741 gtcataatca aggacctcca gctgcttgga ctcgtggcgg cgctgctcat ggccgatgtc
     1801 atactcttga tgacgtgggt gttgaccgac cctatccagt gtcttcaaat tctgagcgtg
     1861 tcaatgacag tcaccggcaa ggatgtctcc tgtacatcca cctctacaca tttttgtgcc
     1921 tcacgataca gcgacgtgtg gattgccctc atctgggggt gcaaagggtt gctcctgctt
     1981 tatggcgcct atctcgccgg gctgaccggc catgttagca gcccccctgt aaatcagagc
     2041 cttacaatta tggtcggggt taacctgctt gtgctggctg ccgggcttct tttcgtcgtg
     2101 actcgatatc tccacagttg gcccaacttg gtttttggcc tgaccagcgg cgggatcttc
     2161 gtttgtacta cgaccattaa ctgcttcatc ttcattccac agttgaagca gtggaaagca
     2221 tttgaggaag aaaaccagac tataagaaga atggccaagt acttctcaac accaaataaa
     2281 agtttccata cccagtacgg agaggaggag aattgccacc cacgcgggga gaaaagcagc
     2341 atggagcgat tgctgaccga gaaaaacgcc gtcatagagt ccttgcagga gcaggtgaac
     2401 aacgccaaag aaaagatagt tcggctcatg tccgctgaat gcacttatga tctgcctgag
     2461 ggggccgcgc cgcccgcttc atccccgaac aaggatgtgc aggcagtagc ttctgtgcac
     2521 actttggccg ccgcacaggg gccgtctggg cacctgtcag actttcagaa cgatcccggc
     2581 atggctgcaa gggacagcca gtgcacaagt ggacccagta gctatgccca gagtcttgaa
     2641 ggcccaggga aggactccag cttttctcct ggtaaggagg aaaaaataag cgattcaaag
     2701 gattttagcg atcacctcga tagcgggtgc agtcagaaac cctggacaga gcagtccctt
     2761 ggtcccgaac ggggtgacca ggtgcccatg aaccccagtc agtctctgct tccagaaagg
     2821 ggtgggtctg acccacagcg gcagcggcac ctggagaata gcgaggagcc accggagcgc
     2881 cggtctagag taagcagtgt tattcgcgaa aaactgcagg aagtccttca ggaccttgga
     2941 ctgggaccag aagcttccct gtcaaccgcg ccaagctgcc accaacagac ctggaaaaac
     3001 tctgctgcgt tcagtccaca gaagatgccc cttagcaagg agctgggctt ttctccatac
     3061 atggttcggc ggagaagagc cgctcaaaga gctcgcagtc acttccccgg tagtgccccc
     3121 tctagtgtgg gccacagagc caataggaca gtccccggtg ctcactcccg gttgcacgtt
     3181 cagaatgggg acagtcctag tttggcacca caaacaaccg actctagggt ccggcgcccc
     3241 tcatccagaa agccttctct gccaagcgat ccacaagaca gacccggaac tctggagggt
     3301 agcaagcaga gccagaccga gcccgagggc gctcgggggt caaaggccgc cttccttaga
     3361 cagccttctg gctccggtcg cgctcctagt cctgctgctc cttgcctttc caaagcatca
     3421 cccgaccttc ccgaacagtg gcagctgtgg cctcctgtgc cttctggctg cgctagcctt
     3481 tcctcccagc acagttactt tgacaccgag agcagcagta gcgatgagtt cttttgtagg
     3541 tgtcaccgac catattgcga aatatgcttt cagtcctcta gcgactcaag cgattctgga
     3601 acatccgata ctgaccccga acctacagga ggtcttgcta gctgggagaa gctgtgggcc
     3661 cgatccaagc ccatcgtaaa cttcaaagac gatctgaagc ccactctggt aatcgatacc
     3721 ggtggacgca ccggtgaatt cgatgaggcc agcaccaccg tgtctaagac cgagacttct
     3781 caggtggccc ctgcctaact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3841 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3901 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3961 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4021 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4081 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4141 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4201 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4261 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4321 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4381 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4441 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4501 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4561 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4621 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4681 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4741 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4801 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4861 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4921 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4981 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5041 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5101 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5161 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5221 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5281 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5341 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5401 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5461 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5521 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5581 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5641 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5701 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5761 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5821 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5881 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5941 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6001 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6061 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6121 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6181 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6241 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6301 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6361 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6421 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6481 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6541 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6601 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6661 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6901 gtggtggcct aactacggct acactagaag aacggtattt ggtatctgcg ctctgctgaa
     6961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7201 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7261 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7321 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7381 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7441 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7501 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7561 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7621 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7681 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7741 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7801 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7861 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7921 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7981 acccactc
//
