LOCUS       sequence-495861-        8529 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted human GPR124 ECD, N-terminal 3xFLAG 
            tag, C-terminal 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8529)
  AUTHORS   Vallon M, Yuki K, Nguyen TD, Chang J, Yuan J, Siepe D, Miao Y, 
            Essler M, Noda M, Garcia KC, Kuo CJ
  TITLE     A RECK-WNT7 Receptor-Ligand Interaction Enables Isoform-Specific 
            Regulation of Wnt Bioavailability.
  JOURNAL   Cell Rep. 2018 Oct 9;25(2):339-349.e9. doi: 
            10.1016/j.celrep.2018.09.045.
  PUBMED    30304675
REFERENCE   2  (bases 1 to 8529)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2018.09"; journalName: "Cell Rep"; date: 
            "2018-10-9- 9"; volume: "25"; issue: "2"; pages: "339-349.e9"
COMMENT     Sequence Label: p3xFLAG-CMV-9-sGPR124-His
FEATURES             Location/Qualifiers
     source          1..8529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             932..976
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             977..1042
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3242..3259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3270..3892
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3469..3488)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3916..3936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3934..4232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4286..5080
                     /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(4340..4359)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4950..4969
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5732..5866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5769..5788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5823..5842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5871..5890)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5872..5890)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5904..5920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5904..5920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5917..5939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5928..5944
                     /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(5952..5958,5959..5976,5977..5982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5997..6018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6135..6152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6306..6894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6386..6405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7065..7856,7857..7925))
                     /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     7688..7707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7926..8030)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8144..8163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8371..8392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcac catgtctgca cttctgatcc tagctcttgt
      961 tggagctgca gttgctgact acaaagacca tgacggtgat tataaagatc atgacatcga
     1021 ttacaaggat gacgatgaca agcttgcggc cgcgaattca gcgcccggct gcccgctatc
     1081 catccgcagc tgcaagtgct cgggggagcg gcccaagggg ctgagcggcg gcgtccctgg
     1141 cccggctcgg cggagggtgg tgtgcagcgg cggggacctc ccggagcctc ccgagcccgg
     1201 ccttctgcct aacggcaccg ttaccctgct cttgagcaat aacaagatca cggggctccg
     1261 caatggctcc ttcctgggac tgtcactgct ggagaagctg gacctgagga acaacatcat
     1321 cagcacagtg cagccgggcg ccttcctggg cctgggggag ctgaagcgtt tagatctctc
     1381 caacaaccgg attggctgtc tcacctccga gaccttccag ggcctcccca ggcttctccg
     1441 actaaacata tctggaaaca tcttctccag tctgcaacct ggggtctttg atgagctgcc
     1501 agcccttaag gttgtggact tgggcaccga gttcctgacc tgtgactgcc acctgcgctg
     1561 gctgctgccc tgggcccaga atcgctccct gcagctgtcg gaacacacgc tctgtgctta
     1621 ccccagtgcc ctgcatgctc aggccctggg cagcctccag gaggcccagc tctgctgcga
     1681 gggggccctg gagctgcaca cacaccacct catcccgtcc ctacgccaag tggtgttcca
     1741 gggggatcgg ctgcccttcc agtgctctgc cagctacctg ggcaacgaca cccgcatccg
     1801 ctggtaccac aaccgagccc ctgtggaggg tgatgagcag gcgggcatcc tcctggccga
     1861 gagcctcatc cacgactgca ccttcatcac cagtgagctg acgctgtctc acatcggcgt
     1921 gtgggcctca ggcgagtggg agtgcaccgt gtccatggcc caaggcaacg ccagcaagaa
     1981 ggtggagatc gtggtgctgg agacctctgc ctcctactgc cccgccgagc gtgttgccaa
     2041 caaccgcggg gacttcaggt ggccccgaac tctggctggc atcacagcct accagtcctg
     2101 cctgcagtat cccttcacct cagtgcccct gggcgggggt gccccgggca cccgagcctc
     2161 ccgccggtgt gaccgtgccg gccgctggga gccaggggac tactcccact gtctctacac
     2221 caacgacatc accagggtgc tgtacacctt cgtgctgatg cccatcaatg cctccaatgc
     2281 gctgaccctg gctcaccagc tgcgcgtgta cacagccgag gccgctagct tttcagacat
     2341 gatggatgta gtctatgtgg ctcagatgat ccagaaattt ttgggttatg tcgaccagat
     2401 caaagagctg gtagaggtga tggtggacat ggccagcaac ctgatgctgg tggacgagca
     2461 cctgctgtgg ctggcccagc gcgaggacaa ggcctgcagc cgcatcgtgg gtgccctgga
     2521 gcgcattggg ggggccgccc tcagccccca tgcccagcac atctcagtga atgcgaggaa
     2581 cgtggcattg gaggcctacc tcatcaagcc gcacagctac gtgggcctga cctgcacagc
     2641 cttccagagg agggagggag gggtgccggg cacacggcca ggaagccctg gccagaaccc
     2701 cccacctgag cccgagcccc cagctgacca gcagctccgc ttccgctgca ccaccgggag
     2761 gcccaatgtt tctctgtcgt ccttccacat caagaacagc gtggccctgg cctccatcca
     2821 gctgcccccg agtctattct catcccttcc ggctgccctg gctcccccgg tgcccccaga
     2881 ctgcaccctg caactgctcg tcttccgaaa tggccgcctc ttccacagcc acagcaacac
     2941 ctcccgccct ggagctgctg ggcctggcaa gaggcgtggc gtggccaccc ccgtcatctt
     3001 cgcaggaacc agtggctgtg gcgtgggaaa cctgacagag ccagtggccg tttcgctgcg
     3061 gcactgggct gagggagccg aacctgtggc cgcttggtgg agccaggagg ggcccgggga
     3121 ggctgggggc tggacctcgg agggctgcca gctccgctcc agccagccca atgtcagcgc
     3181 cctgcactgc cagcacttgg gcaatgtggc cgtgctcatg gagctggatg gggggggttc
     3241 tcatcatcat catcatcatt aaggatcccg ggtggcatcc ctgtgacccc tccccagtgc
     3301 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     3361 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     3421 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     3481 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     3541 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     3601 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     3661 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     3721 gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag caggaggacg
     3781 tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc
     3841 ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc
     3901 ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4081 tcccgcccct aactccgccc atcccgcctc cgctaactcc gcccagttcc gcccattctc
     4141 cgccccatgg ctgactaatt ttttttattt atgcaggccg aggccgcctc ggcctctgag
     4201 ctattccaga agtagtgagg aggctttttt ggaggaattg atcagcttgg gatctgatca
     4261 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4321 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4381 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4441 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4501 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4561 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4621 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4681 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4741 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4801 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4861 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4921 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4981 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5041 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5101 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5161 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5221 ggggatctca tgctggagtt cttcgcccac cccgggctcg atcccctcgc gagttggttc
     5281 agctgctgcc tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc
     5341 atccaggaaa ccagcagcgg ctatccgcgc atccatgccc ccgaactgca ggagtgggga
     5401 ggcacgatgg ccgctttggt cgacccggac gggacgctcc tgcgcctgat acagaacgaa
     5461 ttgcttgcag gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc actgcgtgga
     5521 tggagcgctg gcgcctgctg cgcgacggcg agctgctcac cacccactcg ccaagctgga
     5581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgccgatca taatcagcca
     5641 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     5701 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5761 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5821 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaatt ccctatagtg
     5881 agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5941 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6001 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6061 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6721 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6961 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7021 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7081 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7141 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7201 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7261 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7321 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7381 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7441 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7501 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7561 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7621 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7681 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7741 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7801 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7861 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8041 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     8101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     8161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     8281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8341 acgttcttta agagtccact attaaagaac gtggactctt gttccaaact ggaacaacac
     8401 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     8461 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     8521 ttacaattt
//
