LOCUS       sequence-430769-        8045 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 8045)
  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 8045)
  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: CD97-DuET
FEATURES             Location/Qualifiers
     source          1..8045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(60..648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(140..159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(819..1610,1611..1679))
                     /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     1442..1461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1680..1784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1859..1878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2050..2429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2430..2633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2584..2604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2678..2697
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2678..2696
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5451..5468)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5457..5681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5727..6155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5814..5833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6024..6045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6164..6184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6169..6498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6349..6484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6411..6430
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6565..7359
                     /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(6619..6638)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7229..7248
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7533..7654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7570..7589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7624..7643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7703..7719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7703..7719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7716..7738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7727..7743
                     /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(7751..7757,7758..7775,7776..7781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7796..7817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7934..7951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
       61 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      481 aactacggct acactagaag aacggtattt ggtatctgcg ctctgctgaa gccagttacc
      541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
      961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1801 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     1861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     1921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacattt
     1981 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     2041 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     2101 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2161 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2221 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2281 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2341 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2461 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2521 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2581 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     2641 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     2701 cccaagctgg ctagcgttta aacttaagct tagcgccacc atgaagacga tcatcgccct
     2761 gagctacatc ttctgcctgg tattcgccga ctacaaggac gatgatgacg ccagcatcga
     2821 tatggggggg cgggtgttcc tggccttttg tgtttggctg acgctccccg gagcggagac
     2881 acaggattca agggggtgtg cacggtggtg tcctcagaac tctagctgcg tgaacgccac
     2941 agcatgccgg tgtaatccag gctttagtag cttctctgag attatcacca cccctactga
     3001 gacttgcgac gatattaacg agtgtgcaac ccctagcaaa gtgtcatgcg ggaagttttc
     3061 tgactgctgg aacacagagg gcagctatga ctgcgtctgt agcccaggct atgagccagt
     3121 gtctggagct aagaccttca agaatgagag tgagaacact tgccaggatg tggacgagtg
     3181 ccaacagaac ccacgactct gtaagagtta tgggacgtgt gtaaacaccc tgggctcata
     3241 cacttgccaa tgccttccgg gatttaaatt cattccagag gatcccaaag tgtgtacaga
     3301 cgtcaatgag tgcacatccg gacagaaccc ttgccattcc agcacgcact gtctgaataa
     3361 tgtcgggtct taccagtgcc ggtgtagacc tggctggcag ccgatccccg gctccccgaa
     3421 tggacccaat aacacagttt gtgaagatgt cgatgagtgt tcctctggac agcaccaatg
     3481 cgattcatct accgtctgtt ttaacaccgt cggatcttac agttgccgct gccgccccgg
     3541 ttggaaacct cggcatggga ttccaaataa ccagaaagac acagtgtgcg aagacatgac
     3601 attcagcaca tggactcctc ccccgggcgt ccatagtcaa acccttagca gattctttga
     3661 caaagtccag gatctgggaa gagatagcaa aacaagctcc gccgaggtga ctatccagaa
     3721 cgttatcaag ctggtagatg agctgatgga ggccccgggc gacgtcgagg cgctcgctcc
     3781 cccggtccgg catctgattg ccacacagct tttgagcaat ctggaggata ttatgcgcat
     3841 actggctaaa agcctgccaa agggcccgtt cacgtatatt agcccctcca acacagagct
     3901 tacgctcatg atccaggaac gaggggataa gaatgtgaca atgggccagt cctccgcgag
     3961 gatgaagctc aactgggcag ttgctgccgg agccgaagac cctggccctg ccgtcgctgg
     4021 aatactgagt atacagaata tgactacact gcttgccaac gctagcctga acctgcattc
     4081 caaaaagcaa gcggaactcg aagagattta cgaaagctcc attagaggcg tgcaacttcg
     4141 caggctgagt gcagtgaact ctatattcct gtctcacaac aataccaaag agctgaattc
     4201 ccctatactc ttcgccttca gccatctgga atcatccgac ggagaggccg ggcgggatcc
     4261 cccagcaaag gacgtgatgc ctggtcctag acaggaactc ttgtgtgcct tctggaagtc
     4321 cgactctgac aggggaggcc actgggctac cgagggctgc caagtgctgg gctccaagaa
     4381 cgggagtact acatgtcagt gtagtcacct ctctagtttc gccatcctta tggctcacta
     4441 cgatgtagaa gattggaaat tgacactgat cacgagggtg ggactcgcct tgtccctgtt
     4501 ctgcctgctc ctgtgcatct tgacattcct cctcgttcgc cccatacaag gatctaggac
     4561 cacaatccac ctgcatttgt gtatctgcct gttcgttgga agcaccatct tccttgccgg
     4621 cattgaaaat gaggggggac aagtgggact gcggtgtaga ctggtggctg gtctcctgca
     4681 ctactgcttc ctggctgcgt tctgctggat gtctctggag ggcctcgaac tgtacttcct
     4741 tgttgttaga gtgttccagg gccagggact ctctacacgg tggctctgcc tgatcggcta
     4801 cggcgtgccc cttctgatcg tcggtgtgag cgccgcgatc tactcaaaag gatacggccg
     4861 cccccgctat tgttggctgg acttcgaaca gggctttctc tggtcctttc tgggtcccgt
     4921 gacattcatt atcttgtgca acgcagttat ctttgtcacc accgtgtgga aattgacaca
     4981 aaaattctcc gagatcaatc ccgatatgaa gaagctgaag aaggccagag ctcttaccat
     5041 cacagcaatt gcgcagctgt ttctgctggg gtgtacctgg gtctttggac tcttcatatt
     5101 tgatgacaga agtttggtgc tgacttatgt gtttactatt ctgaactgcc ttcagggtgc
     5161 cttcctctat cttctccatt gcctgctgaa caagaaagtc cgcgaggaat atcgaaagtg
     5221 ggcctgtctg gtcgccgggg ggagtaagta ctctgagttt acctccacta catctggaac
     5281 gggccacaat cagactagag ctctccgggc cagcgagagt ggaatcatcg ataccggtgg
     5341 acgcaccggt gaattcgatg aggccagcac caccgtgtct aagaccgaga cttctcaggt
     5401 ggcccctgcc taactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     5461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5641 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     5701 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     5761 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5821 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5881 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5941 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6001 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6061 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6121 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     6181 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6241 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6301 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6361 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6421 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6481 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     6541 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6601 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6661 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6721 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6781 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6841 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6901 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6961 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     7021 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     7081 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     7141 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     7201 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     7261 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     7321 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     7381 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7441 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7501 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     7561 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7621 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     7681 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7741 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7801 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7861 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8041 agaac
//
