LOCUS       sequence-495863-        8034 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for human GPR124 lacking the ECD, N-terminal 
            3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8034)
  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 8034)
  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-GPR124 Delta-ECD
FEATURES             Location/Qualifiers
     source          1..8034
                     /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"
     polyA_signal    2775..3397
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3421..3441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3439..3737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3791..4585
                     /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(3845..3864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4455..4474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5237..5371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5274..5293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5328..5347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5376..5395)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5377..5395)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5409..5425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5409..5425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5422..5444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5433..5449
                     /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(5457..5463,5464..5481,5482..5487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5502..5523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5640..5657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5811..6399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5891..5910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6570..7361,7362..7430))
                     /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     7193..7212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7431..7535)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7649..7668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7876..7897
                     /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 cccgtggtat acccctgcac
     1081 ggccttgctg ctgctctgcc tcttcgccac catcatcacc tacatcctca accacagctc
     1141 catccgtgtg tcccggaaag gctggcacat gctgctgaac ttgtgcttcc acatagccat
     1201 gacctctgct gtctttgcgg ggggcatcac actcaccaac taccagatgg tctgccaggc
     1261 ggtgggcatc accctgcact actcctccct atccacgctg ctctggatgg gcgtgaaggc
     1321 gcgagtgctc cataaggagc tcacctggag ggcaccccct ccgcaagaag gggaccccgc
     1381 tctgcctact cccagtccta tgctccggtt ctatttgatc gctggaggga ttccactcat
     1441 tatctgtggc atcacagctg cagtcaacat ccacaactac cgggaccaca gcccctactg
     1501 ctggctggtg tggcgtccaa gccttggcgc cttctacatc cctgtggctt tgattctgct
     1561 catcacctgg atctatttcc tgtgcgccgg gctacgctta cggggtcctc tggcacagaa
     1621 ccccaaggcg ggcaacagca gggcctccct ggaggcaggg gaggagctga ggggttccac
     1681 caggctcagg ggcagcggcc ccctcctgag tgactcaggt tcccttcttg ctactgggag
     1741 cgcgcgagtg gggacgcccg ggcccccgga ggatggtgac agcctctatt ctccgggagt
     1801 ccagctaggg gcgctggtga ccacgcactt cctgtacttg gccatgtggg cctgcggggc
     1861 tctggcagtg tcccagcgct ggctgccccg ggtggtgtgc agctgcttgt acggggtggc
     1921 agcctccgcc ctgggcctct tcgtcttcac tcaccactgt gccaggcgga gggacgtgag
     1981 agcctcgtgg cgcgcctgct gcccccctgc ctctcccgcg gccccccatg ccccgccccg
     2041 ggccctgccc gccgccgcag aggacggttc cccggtgttc ggggaggggc ccccctccct
     2101 caagtcctcc ccaagcggca gcagcggcca tccgctggct ctgggcccct gcaagctcac
     2161 caacctgcag ctggcccaga gtcaggtgtg cgaggcgggg gcggcggccg gcggggaagg
     2221 agagccggag ccggcgggca cccggggaaa cctcgcccac cgccacccca acaacgtgca
     2281 ccacgggcgt cgggcgcaca agagccgggc caagggacac cgcgcggggg aggcctgcgg
     2341 caagaaccgg ctcaaggccc tgcgcggggg cgcggcgggg gcgctggagc tgctgtccag
     2401 cgagagcggt agtctgcaca acagccccac cgacagctac ctgggcagca gccgcaacag
     2461 cccgggcgcc ggcctgcagc tggaaggcga gcccatgctc acgccgtccg agggcagcga
     2521 caccagcgcc gcgccgcttt ctgaggcggg ccgggcaggc cagcgccgca gcgccagccg
     2581 cgacagtctc aagggcggcg gcgcgctgga gaaggagagc catcgccgct cgtacccgct
     2641 caacgccgcc agcctaaacg gcgcccccaa ggggggcaag tacgacgacg tcaccctgat
     2701 gggcgcggag gtagccagcg gcggctgcat gaagaccgga ctctggaaga gcgaaactac
     2761 cgtctaagga tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     2821 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     2881 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     2941 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     3001 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     3061 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     3121 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     3181 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     3241 tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag acacagcata
     3301 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct
     3361 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt
     3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcctccgcta actccgccca gttccgccca ttctccgccc catggctgac
     3661 taattttttt tatttatgca ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3721 tgaggaggct tttttggagg aattgatcag cttgggatct gatcaagaga caggatgagg
     3781 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggctg cttgggtgga
     3841 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3901 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3961 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4021 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4081 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4141 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4201 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4261 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4321 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4381 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4441 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4501 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4561 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4621 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4681 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4741 gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt ggttcagctg ctgcctgagg
     4801 ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg tcggcatcca ggaaaccagc
     4861 agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt ggggaggcac gatggccgct
     4921 ttggtcgacc cggacgggac gctcctgcgc ctgatacaga acgaattgct tgcaggcatc
     4981 tcatgagtgt gtcttcccgt tttccgcctg aggtcactgc gtggatggag cgctggcgcc
     5041 tgctgcgcga cggcgagctg ctcaccaccc actcgccaag ctggaaccgt aaaaaggccg
     5101 cgttgctggc gtttttccat aggctccgcc gatcataatc agccatacca catttgtaga
     5161 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5221 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5341 actcatcaat gtatcttatc atgtctggat caattcccta tagtgagtcg tattaaattc
     5401 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5461 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5521 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5581 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5641 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5701 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5761 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5821 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5881 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5941 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6001 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6061 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6121 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6181 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6241 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6301 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6361 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6421 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6481 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6541 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6601 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6661 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6721 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6781 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6841 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6901 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6961 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7021 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7081 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7141 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7201 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7261 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7321 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7381 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7441 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7501 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7561 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     7621 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     7681 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     7741 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     7801 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaagagt
     7861 ccactattaa agaacgtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     7921 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     7981 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca attt
//
