LOCUS       sequence-430782-        6644 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 6644)
  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 6644)
  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: CXCR3-DuET
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     600..619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(838..942)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1017..1036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1208..1587
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1588..1791
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1742..1762
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1836..1855
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1836..1854
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3208..3225)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3214..3438
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3484..3912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3571..3590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3781..3802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3921..3941)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3926..4255
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4106..4241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4168..4187
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4322..5116
                     /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(4376..4395)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4986..5005
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5290..5411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5327..5346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5381..5400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5460..5476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5460..5476)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5473..5495)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5484..5500
                     /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(5508..5514,5515..5532,5533..5538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5553..5574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5691..5708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5862..6450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5942..5961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6621..6644,1..768,769..837))
                     /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 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
       61 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
      121 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
      181 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
      241 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
      301 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      361 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      421 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      481 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      541 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
      601 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
      661 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
      721 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
      781 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
      841 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
      901 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
      961 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     1021 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     1081 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacatttgc
     1141 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     1201 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1261 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1321 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1381 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1441 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1501 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1561 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     1621 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1681 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     1741 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     1801 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     1861 caagctggct agcgtttaaa cttaagctta gcgccaccat gaagacgatc atcgccctga
     1921 gctacatctt ctgcctggta ttcgccgact acaaggacga tgatgacgcc agcatcgata
     1981 tggttctgga agtttccgac caccaggtgc ttaatgatgc ggaagttgcg gcactgctgg
     2041 aaaactttag ctcatcctat gactacggcg agaacgagtc agactcttgc tgtacctccc
     2101 ctccttgccc ccaagacttt agcttgaatt tcgatcgggc cttcctgccc gcactttact
     2161 ctctgctgtt cctgttggga ctgttgggca acggggccgt ggcagcagta ctgttgagtc
     2221 gaaggactgc cctgtccagt accgatacat ttctgctgca cctcgccgtg gctgacacac
     2281 tgctggttct gaccctgccc ctttgggcag tcgatgcagc tgtccagtgg gttttcggga
     2341 gtggactttg taaggtggca ggggcgctgt tcaatatcaa cttctacgcc ggagcactgc
     2401 tgctggcttg tataagcttc gaccgatatc tgaacattgt gcacgccacg caactctata
     2461 gaagaggccc tccagcaagg gttacactca cctgtcttgc agtttggggg ctgtgcttgc
     2521 ttttcgcatt gcccgacttc attttcctga gtgctcacca tgacgagcgg ctgaacgcca
     2581 ctcattgtca atacaacttc cctcaggtgg ggagaacagc gctgcgagtg ctgcagttgg
     2641 tggccggatt cctgctgcct ttgctggtga tggcgtactg ctacgcgcat attttggcgg
     2701 tcttgttggt ttctagaggg caaaggcggc tgagggctat gcgactggtc gttgtggtgg
     2761 ttgtcgcctt cgcgctctgc tggaccccat accacctggt ggtgctggtt gatatactga
     2821 tggatttggg agccctggcg aggaattgcg gtagagaaag ccgggtcgac gttgcaaaaa
     2881 gcgtcacctc agggctgggc tacatgcact gttgtctgaa cccccttctg tacgcattcg
     2941 tgggggtgaa gttccgggaa cgcatgtgga tgttgttgct gcgactgggt tgccctaatc
     3001 agcgcggtct gcagcgacag cccagttctt ctcgaaggga ctcatcatgg tccgaaacct
     3061 cagaggctag ttattctggc ctcatcgata ccggtggacg caccggtgaa ttcgatgagg
     3121 ccagcaccac cgtgtctaag accgagactt ctcaggtggc ccctgcctaa ctcgagtcta
     3181 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3241 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3301 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3361 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3421 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3481 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3841 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3901 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3961 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4081 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4141 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4201 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4261 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4321 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4381 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4441 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4501 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4561 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4621 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4681 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4741 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4801 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4861 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4921 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4981 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5041 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5101 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5161 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5221 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5281 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5401 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6301 agaacggtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggc
//
