LOCUS       sequence-430809-        6545 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 6545)
  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 6545)
  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: GPR119-DuET
FEATURES             Location/Qualifiers
     source          1..6545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     509..528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(747..851)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(926..945)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1117..1496
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1497..1700
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1651..1671
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1745..1764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1745..1763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3018..3035)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3024..3248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3294..3722
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3591..3612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3731..3751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3736..4065
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3916..4051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3978..3997
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4132..4926
                     /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(4186..4205)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4796..4815
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5100..5221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5137..5156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5191..5210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5270..5286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5270..5286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5283..5305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5294..5310
                     /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(5318..5324,5325..5342,5343..5348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5363..5384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5501..5518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5672..6260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5752..5771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6431..6545,1..677,678..746))
                     /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 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
       61 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
      121 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
      181 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
      241 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
      301 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
      361 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
      421 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
      481 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
      541 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
      601 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
      661 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
      721 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
      781 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
      841 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
      901 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      961 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     1021 caacaaggca aggcttgacc gacatttgca tgaagaatct gcttagggtt aggcgttttg
     1081 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
     1141 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1201 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     1261 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     1321 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     1381 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1441 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1501 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1561 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1621 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     1681 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
     1741 aaattaatac gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttag
     1801 cgccaccatg aagacgatca tcgccctgag ctacatcttc tgcctggtat tcgccgacta
     1861 caaggacgat gatgacgcca gcatcgatat ggagtcctct tttagctttg gggtgatact
     1921 ggccgtcctt gcatctctca tcattgcaac caacactctg gtggcagtgg cggtgctcct
     1981 gttgatccac aagaatgacg gagtaagcct gtgctttaca ctcaacctcg ctgttgccga
     2041 tacattgatc ggtgttgcta tctcagggct gctgactgac cagctgtcat ccccaagcag
     2101 accaacacag aagacgctgt gctccctgcg catggctttc gtgacctcta gtgccgctgc
     2161 cagcgtgttg acggttatgc tgattacttt cgataggtac ctggcaatca agcaaccttt
     2221 tcggtatctg aaaataatgt ctggatttgt cgccggcgca tgtatcgccg gcctctggct
     2281 ggtcagctac ctcattggat tcctccctct gggcattccc atgttccagc agacagccta
     2341 taaggggcaa tgtagtttct ttgcggtttt tcacccacac ttcgtgctca ctctctcatg
     2401 cgtgggtttc tttccagcga tgttgctttt tgtgttcttc tactgtgata tgctgaagat
     2461 tgcgtccatg catagccagc aaatacgcaa aatggagcat gcaggcgcca tggcgggggg
     2521 gtatcggagc cctagaaccc cgtccgactt taaggcactg cggactgtat cagttctgat
     2581 tggaagtttt gccctttctt ggaccccttt tcttatcacc ggcattgtgc aggttgcctg
     2641 ccaggaatgc cacctctatc tcgtgctgga acggtatctg tggcttctcg gagtcggaaa
     2701 ctctctcctc aatcctctga tctacgcgta ctggcaaaag gaggtacggc tccagcttta
     2761 ccacatggcg ctcggggtca agaaggttct gacaagtttc ctgctgttct tgtcagcccg
     2821 caattgtggg ccagaacggc cccgggagtc ctcctgccac atagttacta tttcctccag
     2881 cgaatttgat ggaatcgata ccggtggacg caccggtgaa ttcgatgagg ccagcaccac
     2941 cgtgtctaag accgagactt ctcaggtggc ccctgcctaa ctcgagtcta gagggcccgt
     3001 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3061 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3121 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3181 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3241 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3301 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3361 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3421 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3481 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3541 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3601 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3661 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3721 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3841 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3901 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3961 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4021 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4081 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4141 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4201 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4261 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4321 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4381 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4441 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4501 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4561 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4621 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4681 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4741 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4801 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4861 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4921 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4981 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5041 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5101 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5161 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5221 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5281 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5341 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5401 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5461 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5521 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacggtat
     6121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6421 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6481 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6541 catct
//
