LOCUS       sequence-430821-        6785 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 6785)
  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 6785)
  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: GPR19-DuET
FEATURES             Location/Qualifiers
     source          1..6785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     375..394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(613..717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(792..811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        983..1362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1363..1566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1517..1537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1611..1630
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1611..1629
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3124..3141)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3130..3354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3400..3828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3487..3506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3697..3718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3837..3857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3842..4171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4022..4157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4084..4103
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4238..5032
                     /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(4292..4311)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4902..4921
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5206..5327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5243..5262)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5297..5316
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5376..5392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5376..5392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5389..5411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5400..5416
                     /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(5424..5430,5431..5448,5449..5454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5469..5490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5607..5624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5778..6366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5858..5877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6537..6785,1..543,544..612))
                     /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"
ORIGIN
        1 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
       61 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      121 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      181 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      241 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      301 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      361 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      421 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      481 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      541 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      601 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      661 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      721 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      781 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      841 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      901 ttgaccgaca tttgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      961 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     1021 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1081 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1141 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1201 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1261 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1321 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1381 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1441 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1501 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1561 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     1621 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttagcgcc accatgaaga
     1681 cgatcatcgc cctgagctac atcttctgcc tggtattcgc cgactacaag gacgatgatg
     1741 acgccagcat cgatatggta ttcgctcatc gcatggacaa cagcaagcct catctgatca
     1801 ttcctactct cctggtgccg ttgcaaaaca ggagctgtac cgagaccgcc actccacttc
     1861 cgtcccaata cctgatggaa ttgagcgaag aacacagctg gatgtctaat caaacggacc
     1921 tccactatgt gctgaaaccg ggagaggttg ccaccgccag tatcttcttt ggtattcttt
     1981 ggctttttag catattcgga aacagcctcg tgtgccttgt gattcatcgc tctcgccgga
     2041 cacaaagcac cacaaactac ttcgtggtct ccatggcatg tgcagacttg ctgatctcag
     2101 tggcctccac cccgttcgtg cttctgcagt tcacaacagg ccggtggacc cttggctctg
     2161 ctacctgtaa agttgtgaga tactttcagt acctcacgcc tggcgtgcaa atatacgttc
     2221 tcttgagcat ttgtatagat cggttctata ctatcgtgta tcccctgtca tttaaagtca
     2281 gtcgcgaaaa ggccaaaaaa atgatagcgg ctagctggat ctttgatgct gggtttgtta
     2341 ctccagttct gttcttttac gggagcaact gggattccca ttgcaactac tttcttccga
     2401 gcagctggga agggaccgcc tacaccgtga tacacttcct tgtcggattt gtcattccgt
     2461 ctgtccttat catattgttc taccagaagg tgatcaagta tatctggcga atcggcacgg
     2521 atggccgaac agtgcggaga acgatgaaca ttgttcccag aaccaaagtt aagacaataa
     2581 aaatgttcct catccttaat ctcctgttcc tgctgtcctg gttgcccttt cacgtagctc
     2641 agttgtggca cccccacgaa caggactaca aaaaatcatc tctggtattc acggccatca
     2701 catggatctc cttctcttct agcgcttcaa aaccgactct gtatagtatt tataacgcta
     2761 actttaggag aggcatgaag gagaccttct gtatgtcctc catgaagtgc tatagatcta
     2821 acgcctacac aataactact tcatccagga tggcaaaaaa gaactacgtc gggatctccg
     2881 agatccccag catggctaag actatcacta aggactccat ctacgactcc ttcgatagag
     2941 aagccaagga gaagaagctg gcttggccta tcaactcaaa cccacccaac actttcgtga
     3001 tcgataccgg tggacgcacc ggtgaattcg atgaggccag caccaccgtg tctaagaccg
     3061 agacttctca ggtggcccct gcctaactcg agtctagagg gcccgtttaa acccgctgat
     3121 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3181 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3241 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3301 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3361 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3421 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3481 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3541 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3601 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3661 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3721 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3781 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3841 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3901 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3961 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4021 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4081 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4141 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4201 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4261 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4321 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4381 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4441 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4501 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4561 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4621 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4681 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4741 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4801 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4861 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4921 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4981 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5041 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5101 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5161 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5221 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5281 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5341 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5401 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5461 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5521 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5581 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5641 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5701 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cggtatttgg tatctgcgct
     6241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6781 attaa
//
