LOCUS       sequence-430780-        6590 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 6590)
  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 6590)
  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: CXCR1-DuET
FEATURES             Location/Qualifiers
     source          1..6590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(36..827,828..896))
                     /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     659..678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(897..1001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1076..1095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1267..1646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1647..1850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1801..1821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1895..1914
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1895..1913
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3213..3230)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3219..3443
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3489..3917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3576..3595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3786..3807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3926..3946)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3931..4260
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4111..4246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4173..4192
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4327..5121
                     /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(4381..4400)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4991..5010
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5295..5416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5332..5351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5386..5405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5465..5481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5465..5481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5478..5500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5489..5505
                     /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(5513..5519,5520..5537,5538..5543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5558..5579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5696..5713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5867..6455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5947..5966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
       61 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
      361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
      481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
      541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
      601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
      661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
      721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
      781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
      841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1021 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     1081 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     1141 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacatttgca
     1201 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     1261 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     1321 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     1381 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     1441 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     1501 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     1561 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     1621 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     1681 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     1741 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1801 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
     1861 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
     1921 aagctggcta gcgtttaaac ttaagcttag cgccaccatg aagacgatca tcgccctgag
     1981 ctacatcttc tgcctggtat tcgccgacta caaggacgat gatgacgcca gcatcgatat
     2041 gtccaatatc accgatcccc agatgtggga tttcgatgac ctcaacttca cgggcatgcc
     2101 tcccgctgat gaagattaca gcccatgcat gctggagaca gagaccctga acaaatacgt
     2161 agttataatc gcatacgctc tggtcttcct tctgtctctc ctgggaaata gtctggttat
     2221 gcttgttata ctgtacagca gagtgggccg ctctgttact gacgtctacc tcctgaacct
     2281 ggcactcgct gacttgttgt ttgctttgac cctgccgatc tgggccgcct ccaaggtaaa
     2341 cgggtggatc tttggcacat tcttgtgcaa ggtagtgtca ttgctgaaag aggtgaactt
     2401 ttactccggg atactgttgc tggcttgcat tagcgtcgac aggtacctcg caatagtgca
     2461 cgctactcgc acactcactc aaaagcggca cctggtcaag tttgtgtgcc tcgggtgctg
     2521 ggggctctct atgaacttgt ccctcccctt ttttctgttc agacaggcat atcaccccaa
     2581 caattcctct cctgtctgct atgaggtact cggcaacgat actgcaaagt ggaggatggt
     2641 actgcggatt ttgcctcaca ccttcggctt tatcgtgcct ctgttcgtaa tgctgttctg
     2701 ctacggattt acgcttcgca ctctgttcaa ggcacacatg gggcagaagc acagggctat
     2761 gcgggtgatc tttgccgtag tgctgatctt cctgctctgt tggctcccgt ataatctggt
     2821 cctccttgca gataccctga tgcggactca ggtcatccag gaatcatgtg aacgccgcaa
     2881 taatattggc cgagcccttg atgctacaga gattttgggc tttctgcaca gctgcctcaa
     2941 cccaatcatc tatgcgttta ttggccaaaa tttccgccac ggatttctca aaattctcgc
     3001 aatgcacggc ctggtgtcca aggagtttct ggctagacac cgggtgacaa gttatacaag
     3061 cagttcagtt aatgtctctt caaatttgat cgataccggt ggacgcaccg gtgaattcga
     3121 tgaggccagc accaccgtgt ctaagaccga gacttctcag gtggcccctg cctaactcga
     3181 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3241 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3301 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3361 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3421 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3481 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3541 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3601 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3661 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3721 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3781 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3841 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3901 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4141 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4201 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4261 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4321 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4381 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4441 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4501 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4561 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4621 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4681 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4741 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4801 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4861 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4921 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4981 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5041 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5101 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5161 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5221 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5281 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5341 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5401 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5461 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5521 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5581 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5641 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5701 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6301 ctagaagaac ggtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
//
