LOCUS       sequence-430736-        6542 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 6542)
  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 6542)
  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: SUCNR1-DuET
FEATURES             Location/Qualifiers
     source          1..6542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      229..657
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(316..335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     526..547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(666..686)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        671..1000
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      851..986
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     913..932
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1067..1861
                     /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(1121..1140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1731..1750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2035..2156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2072..2091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2126..2145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2205..2221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2218..2240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2229..2245
                     /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(2253..2259,2260..2277,2278..2283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2298..2319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2436..2453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2607..3195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2687..2706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3366..4157,4158..4226))
                     /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     3989..4008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4227..4331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4406..4425)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4597..4976
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4977..5180
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5131..5151
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5225..5244
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5225..5243
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6495..6512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(6501..6542,1..183)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
       61 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
      121 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
      181 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
      241 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      301 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      361 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      421 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
      481 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      541 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
      601 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
      661 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
      721 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
      781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
      841 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
      901 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
      961 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     1021 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     1081 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     1141 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     1201 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     1261 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     1321 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     1381 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     1441 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     1501 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     1561 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     1621 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     1681 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     1741 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     1801 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     1861 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     1921 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     1981 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     2041 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2101 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2161 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     2221 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2281 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2341 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2401 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2461 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2521 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2581 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2641 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2701 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2761 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2821 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2881 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2941 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3001 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac ggtatttggt
     3061 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3121 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3181 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3241 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3301 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3361 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3421 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3481 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3541 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3601 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3661 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3721 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3781 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3841 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3901 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3961 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4021 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4081 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4141 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4201 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4261 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4321 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     4381 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     4441 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     4501 caacaaggca aggcttgacc gacatttgca tgaagaatct gcttagggtt aggcgttttg
     4561 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
     4621 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4681 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4741 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4801 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4861 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4921 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     4981 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     5041 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     5101 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     5161 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
     5221 aaattaatac gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttag
     5281 cgccaccatg aagacgatca tcgccctgag ctacatcttc tgcctggtat tcgccgacta
     5341 caaggacgat gatgacgcca gcatcgatat gctgggaatt atggcctgga atgccacctg
     5401 taagaattgg ctggctgcag aggctgccct ggaaaaatat tacctgtcca ttttctatgg
     5461 gatcgagttt gtagtgggtg ttctgggtaa tactattgtg gtgtacggat acatcttctc
     5521 cctgaaaaat tggaattctt ctaatattta tctgttcaat ctttccgtgt ccgatctggc
     5581 tttcctgtgc accttgccga tgctcatcag atcatatgcc aatgggaatt ggatctatgg
     5641 cgacgtcctc tgtatctcca atagatacgt cctgcacgcc aacctgtata cctcaatttt
     5701 gttcttgacg ttcataagta tcgaccgcta cctcatcata aagtacccct ttcgcgaaca
     5761 cctgcttcag aaaaaagaat ttgccattct cattagcttg gctatctggg ttcttgtgac
     5821 ccttgaactc ctcccgattc tgcccctcat caaccccgtg atcacagata atggcactac
     5881 ctgtaacgac tttgcttcca gtggggaccc gaattacaat ctgatctaca gtatgtgtct
     5941 cacactcctc ggatttctga ttcctctgtt cgtgatgtgt ttcttttatt acaagatagc
     6001 tctgtttttg aagcagagaa acagacaggt ggctaccgct ctcccactgg agaagccact
     6061 gaacctggtc atcatggcgg ttgtgatctt ctctgttctt tttactccat accacgtgat
     6121 gagaaacgtc aggatagcca gtcggctggg ctcatggaag cagtaccaat gcacccaggt
     6181 ggtgataaac agcttctata ttgttactcg gccactggct ttcctgaatt ctgttatcaa
     6241 ccctgtgttc tattttctgc tgggcgacca ctttagggat atgctgatga accagttgcg
     6301 gcacaatttt aagtccctga catccttttc ccggtgggcc cacgaactgc tcctgtcttt
     6361 tagggagaag atcgataccg gtggacgcac cggtgaattc gatgaggcca gcaccaccgt
     6421 gtctaagacc gagacttctc aggtggcccc tgcctaactc gagtctagag ggcccgttta
     6481 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6541 cc
//
