LOCUS       sequence-495870-        8295 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted human GPR124 ECD lacking the HRM 
            domain, N-terminal 3xFLAG tag, C-terminal 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8295)
  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 8295)
  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 Delta-HRM-His
FEATURES             Location/Qualifiers
     source          1..8295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1280..1297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1308..1930
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1507..1526)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1954..1974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1972..2270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2324..3118
                     /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(2378..2397)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2988..3007
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3770..3904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3807..3826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3861..3880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3909..3928)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3910..3928)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3942..3958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3942..3958)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3955..3977)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3966..3982
                     /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(3990..3996,3997..4014,4015..4020))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4035..4056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4173..4190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4344..4932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4424..4443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5103..5894,5895..5963))
                     /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     5726..5745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5964..6068)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6182..6201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6409..6430
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6693..6715
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6707..6724
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6708..6724
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6885..7264
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7265..7468
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7419..7439
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             7499..7543
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             7544..7609
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 agtggtgttc cagggggatc ggctgccctt ccagtgctct gccagctacc tgggcaacga
       61 cacccgcatc cgctggtacc acaaccgagc ccctgtggag ggtgatgagc aggcgggcat
      121 cctcctggcc gagagcctca tccacgactg caccttcatc accagtgagc tgacgctgtc
      181 tcacatcggc gtgtgggcct caggcgagtg ggagtgcacc gtgtccatgg cccaaggcaa
      241 cgccagcaag aaggtggaga tcgtggtgct ggagaccctg tacaccttcg tgctgatgcc
      301 catcaatgcc tccaatgcgc tgaccctggc tcaccagctg cgcgtgtaca cagccgaggc
      361 cgctagcttt tcagacatga tggatgtagt ctatgtggct cagatgatcc agaaattttt
      421 gggttatgtc gaccagatca aagagctggt agaggtgatg gtggacatgg ccagcaacct
      481 gatgctggtg gacgagcacc tgctgtggct ggcccagcgc gaggacaagg cctgcagccg
      541 catcgtgggt gccctggagc gcattggggg ggccgccctc agcccccatg cccagcacat
      601 ctcagtgaat gcgaggaacg tggcattgga ggcctacctc atcaagccgc acagctacgt
      661 gggcctgacc tgcacagcct tccagaggag ggagggaggg gtgccgggca cacggccagg
      721 aagccctggc cagaaccccc cacctgagcc cgagccccca gctgaccagc agctccgctt
      781 ccgctgcacc accgggaggc ccaatgtttc tctgtcgtcc ttccacatca agaacagcgt
      841 ggccctggcc tccatccagc tgcccccgag tctattctca tcccttccgg ctgccctggc
      901 tcccccggtg cccccagact gcaccctgca actgctcgtc ttccgaaatg gccgcctctt
      961 ccacagccac agcaacacct cccgccctgg agctgctggg cctggcaaga ggcgtggcgt
     1021 ggccaccccc gtcatcttcg caggaaccag tggctgtggc gtgggaaacc tgacagagcc
     1081 agtggccgtt tcgctgcggc actgggctga gggagccgaa cctgtggccg cttggtggag
     1141 ccaggagggg cccggggagg ctgggggctg gacctcggag ggctgccagc tccgctccag
     1201 ccagcccaat gtcagcgccc tgcactgcca gcacttgggc aatgtggccg tgctcatgga
     1261 gctggatggg gggggttctc atcatcatca tcatcattaa ggatcccggg tggcatccct
     1321 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     1381 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     1441 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     1501 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     1561 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     1621 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     1681 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     1741 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     1801 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     1861 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     1921 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     1981 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2041 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2101 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgcctccg ctaactccgc
     2161 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcaggccgag
     2221 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat
     2281 cagcttggga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2341 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2401 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     2461 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     2521 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     2581 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     2641 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     2701 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     2761 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     2821 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     2881 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     2941 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3001 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3061 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3121 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3181 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3241 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat
     3301 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg
     3361 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc
     3421 gaactgcagg agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg
     3481 cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc
     3541 ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca
     3601 cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3661 gccgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3721 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3901 gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg
     3961 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4021 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4081 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4141 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4201 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4741 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5041 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5101 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5161 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5221 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5281 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5341 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5401 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5461 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5521 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5581 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5641 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5701 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5761 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5821 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5881 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5941 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6001 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6061 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     6121 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     6181 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6241 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6301 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6361 cctttgacgt tggagtccac gttctttaag agtccactat taaagaacgt ggactcttgt
     6421 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6481 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6541 ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga
     6601 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6661 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     6721 cagtgccaag ctgatctata cattgaatca atattggcaa ttagccatat tagtcattgg
     6781 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     6841 tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac
     6901 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6961 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     7021 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     7081 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     7141 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     7201 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     7261 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     7321 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     7381 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     7441 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagaatt aattcaccat
     7501 gtctgcactt ctgatcctag ctcttgttgg agctgcagtt gctgactaca aagaccatga
     7561 cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagc ttgcggccgc
     7621 gaattcagcg cccggctgcc cgctatccat ccgcagctgc aagtgctcgg gggagcggcc
     7681 caaggggctg agcggcggcg tccctggccc ggctcggcgg agggtggtgt gcagcggcgg
     7741 ggacctcccg gagcctcccg agcccggcct tctgcctaac ggcaccgtta ccctgctctt
     7801 gagcaataac aagatcacgg ggctccgcaa tggctccttc ctgggactgt cactgctgga
     7861 gaagctggac ctgaggaaca acatcatcag cacagtgcag ccgggcgcct tcctgggcct
     7921 gggggagctg aagcgtttag atctctccaa caaccggatt ggctgtctca cctccgagac
     7981 cttccagggc ctccccaggc ttctccgact aaacatatct ggaaacatct tctccagtct
     8041 gcaacctggg gtctttgatg agctgccagc ccttaaggtt gtggacttgg gcaccgagtt
     8101 cctgacctgt gactgccacc tgcgctggct gctgccctgg gcccagaatc gctccctgca
     8161 gctgtcggaa cacacgctct gtgcttaccc cagtgccctg catgctcagg ccctgggcag
     8221 cctccaggag gcccagctct gctgcgaggg ggccctggag ctgcacacac accacctcat
     8281 cccgtcccta cgcca
//
