LOCUS       sequence-430786-        6626 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 6626)
  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 6626)
  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: CXCR7-DuET
FEATURES             Location/Qualifiers
     source          1..6626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      98..686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     587..606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     840..857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    974..995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1010..1015,1016..1033,1034..1040)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1048..1064
                     /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)."
     primer_bind     1053..1075
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1072..1088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1072..1088
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1137..1258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1148..1167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1202..1221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1432..2226)
                     /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(1543..1562)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2153..2172
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2293..2622)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2307..2442
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2361..2380)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2607..2627
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2636..3064)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2746..2767)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2958..2977
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3110..3334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3323..3340
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(4675..4694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4676..4694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4739..4942)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4768..4788)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4943..5322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5494..5513
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5588..5692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5693..5761,5762..6553)
                     /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     complement(5911..5930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
       61 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      121 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      181 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      241 ccgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      301 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      361 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      421 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      481 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      541 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      601 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      661 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      721 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      781 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
      841 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      901 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      961 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     1021 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     1081 gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag
     1141 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1201 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     1261 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     1321 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     1381 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     1441 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     1501 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     1561 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     1621 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     1681 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     1741 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     1801 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     1861 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     1921 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     1981 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     2041 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     2101 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     2161 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     2221 aatcatgcga aacgatcctc atcctgtctc ttgatcagat ccgaaaatgg atatacaagc
     2281 tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     2341 tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     2401 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     2461 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     2521 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     2581 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gaattaattc
     2641 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2701 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2761 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2821 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2881 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2941 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3001 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3061 gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc catagagccc
     3121 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     3181 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     3241 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     3301 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg
     3361 ggccctctag actcgagtta ggcaggggcc acctgagaag tctcggtctt agacacggtg
     3421 gtgctggcct catcgaattc accggtgcgt ccaccggtat cgattttggt agattgttca
     3481 agtgcgctat actcggtttc actaacgcgg gatgcgtcaa tcagcttagt aagcccggtc
     3541 ttggctgagt atttgaatat gaacgctttc atcaactcat acctataatt tcgattaata
     3601 aagctataca ggacgggatt aacgcagcaa tgcacgagag acaaacactg agtcacgtgc
     3661 agtgctgtaa aaagggcatg ttccaggcgg caggtaaatg ggatataatg gagtatggaa
     3721 aagatatcca gcagcactgc aacgtggtat ggcagccagc acaccagaaa aaccacgacg
     3781 tagctaaata taatttttct ggaactatgt ttctcctggt cgctgctcgc tgaaatggcc
     3841 cgcgccaaga ggaagtagaa gaccgcgata attgaaaacg gcacggcgaa gcccagcacc
     3901 acggacacaa gctccatgcc gatgagccac tccttgatgc tatgctcagg gtagaaggag
     3961 cgacagtacg tctcattgtt actggcagaa gttacggttt tcaggtagta tgtgtctggc
     4021 aaggaaacac aaaaggcaag cagccacacg agtatgcaga ccaccctccg caccatcttc
     4081 ttccgggagg agggggtatt ggtgaagtac gtgattgaca ggtacctgtc cactgacatg
     4141 catgtaagga agaagatgga gccaaaaagg ttgatagaaa agatcaggtg ggtcacctta
     4201 caggtaagct cgcccattgg ccattggttg tgttgaacca ggctcaccac ccacactggg
     4261 atagtaagca ccacccaaag atcggcaatg gccaggttga ggatatagca gtgggtgtcg
     4321 tatcctgtag tcttggcctg gatgttcacc catacgacga cactatttgc aatcatgcca
     4381 ataacgaata taaagatgta gatgaagctc agtgtgtaaa gcagaactga cttatttggc
     4441 atattaggac acataactgt gtcgaccaca atgcaatcgc tgctattgca aggccaagag
     4501 atgtctgaaa aatttccagg ctctgaataa tcgaacagat gcaaatccat atcgatgctg
     4561 gcgtcatcat cgtccttgta gtcggcgaat accaggcaga agatgtagct cagggcgatg
     4621 atcgtcttca tggtggcgct aagcttaagt ttaaacgcta gccagcttgg gtctccctat
     4681 agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag
     4741 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     4801 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     4861 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     4921 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     4981 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     5041 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     5101 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     5161 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     5221 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     5281 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc
     5341 gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg caaatgtcgg
     5401 tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca
     5461 cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag
     5521 tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg
     5581 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5641 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5701 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5761 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5821 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5881 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5941 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6001 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6061 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6121 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6181 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6241 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6301 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6361 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6421 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6481 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6541 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6601 tcatttttaa tttaaaagga tctagg
//
