LOCUS       sequence-495741-        7914 bp    DNA     circular SYN 18-DEC-2025
DEFINITION  Expression vector for secreted human GPR124 ECD lacking LRR 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 7914)
  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 7914)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 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-LRR-His
FEATURES             Location/Qualifiers
     source          1..7914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(62..79)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(233..821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(313..332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(992..1783,1784..1852))
                     /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     1615..1634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1853..1957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2071..2090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2298..2319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2582..2604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2596..2613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2597..2613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2774..3153
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3154..3357
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3308..3328
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3388..3432
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             3433..3498
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5083..5100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5111..5733
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5310..5329)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5757..5777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5775..6073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6127..6921
                     /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(6181..6200)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6791..6810
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7573..7707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7610..7629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7664..7683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7712..7731)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7713..7731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7745..7761)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7745..7761)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7758..7780)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7769..7785
                     /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(7793..7799,7800..7817,7818..7823))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7838..7859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
       61 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      121 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      181 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      241 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      301 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      361 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      421 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      481 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      541 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      601 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      661 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      721 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      781 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
      841 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
      901 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
      961 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1021 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1081 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1141 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1201 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1261 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1321 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1381 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1441 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1501 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1561 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     1621 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     1681 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1741 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     1801 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     1861 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1921 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1981 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2041 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2101 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2161 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2221 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaaga
     2281 gtccactatt aaagaacgtg gactcttgtt ccaaactgga acaacactca accctatctc
     2341 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2401 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat
     2461 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2521 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2581 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac attgaatcaa
     2641 tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg
     2701 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
     2761 tgaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     2821 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2881 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2941 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     3001 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
     3061 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
     3121 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat
     3181 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     3241 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
     3301 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     3361 ttagtgaacc gtcagaatta attcaccatg tctgcacttc tgatcctagc tcttgttgga
     3421 gctgcagttg ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     3481 aaggatgacg atgacaagct tgcggccgcg aattcacacc acctcatccc ggccgcgaat
     3541 tcacaccacc tcatcccgtc cctacgccaa gtggtgttcc agggggatcg gctgcccttc
     3601 cagtgctctg ccagctacct gggcaacgac acccgcatcc gctggtacca caaccgagcc
     3661 cctgtggagg gtgatgagca ggcgggcatc ctcctggccg agagcctcat ccacgactgc
     3721 accttcatca ccagtgagct gacgctgtct cacatcggcg tgtgggcctc aggcgagtgg
     3781 gagtgcaccg tgtccatggc ccaaggcaac gccagcaaga aggtggagat cgtggtgctg
     3841 gagacctctg cctcctactg ccccgccgag cgtgttgcca acaaccgcgg ggacttcagg
     3901 tggccccgaa ctctggctgg catcacagcc taccagtcct gcctgcagta tcccttcacc
     3961 tcagtgcccc tgggcggggg tgccccgggc acccgagcct cccgccggtg tgaccgtgcc
     4021 ggccgctggg agccagggga ctactcccac tgtctctaca ccaacgacat caccagggtg
     4081 ctgtacacct tcgtgctgat gcccatcaat gcctccaatg cgctgaccct ggctcaccag
     4141 ctgcgcgtgt acacagccga ggccgctagc ttttcagaca tgatggatgt agtctatgtg
     4201 gctcagatga tccagaaatt tttgggttat gtcgaccaga tcaaagagct ggtagaggtg
     4261 atggtggaca tggccagcaa cctgatgctg gtggacgagc acctgctgtg gctggcccag
     4321 cgcgaggaca aggcctgcag ccgcatcgtg ggtgccctgg agcgcattgg gggggccgcc
     4381 ctcagccccc atgcccagca catctcagtg aatgcgagga acgtggcatt ggaggcctac
     4441 ctcatcaagc cgcacagcta cgtgggcctg acctgcacag ccttccagag gagggaggga
     4501 ggggtgccgg gcacacggcc aggaagccct ggccagaacc ccccacctga gcccgagccc
     4561 ccagctgacc agcagctccg cttccgctgc accaccggga ggcccaatgt ttctctgtcg
     4621 tccttccaca tcaagaacag cgtggccctg gcctccatcc agctgccccc gagtctattc
     4681 tcatcccttc cggctgccct ggctcccccg gtgcccccag actgcaccct gcaactgctc
     4741 gtcttccgaa atggccgcct cttccacagc cacagcaaca cctcccgccc tggagctgct
     4801 gggcctggca agaggcgtgg cgtggccacc cccgtcatct tcgcaggaac cagtggctgt
     4861 ggcgtgggaa acctgacaga gccagtggcc gtttcgctgc ggcactgggc tgagggagcc
     4921 gaacctgtgg ccgcttggtg gagccaggag gggcccgggg aggctggggg ctggacctcg
     4981 gagggctgcc agctccgctc cagccagccc aatgtcagcg ccctgcactg ccagcacttg
     5041 ggcaatgtgg ccgtgctcat ggagctggat ggggggggtt ctcatcatca tcatcatcat
     5101 taaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     5161 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     5221 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     5281 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     5341 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     5401 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     5461 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     5521 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     5581 tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct
     5641 acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg
     5701 cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa
     5761 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5821 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5881 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5941 catcccgcct ccgctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6001 tttttttatt tatgcaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6061 gaggcttttt tggaggaatt gatcagcttg ggatctgatc aagagacagg atgaggatcg
     6121 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6181 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6241 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6301 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6361 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6421 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6481 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6541 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6601 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6661 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6721 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6781 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6841 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6901 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6961 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     7021 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     7081 tcttcgccca ccccgggctc gatcccctcg cgagttggtt cagctgctgc ctgaggctgg
     7141 acgacctcgc ggagttctac cggcagtgca aatccgtcgg catccaggaa accagcagcg
     7201 gctatccgcg catccatgcc cccgaactgc aggagtgggg aggcacgatg gccgctttgg
     7261 tcgacccgga cgggacgctc ctgcgcctga tacagaacga attgcttgca ggcatctcat
     7321 gagtgtgtct tcccgttttc cgcctgaggt cactgcgtgg atggagcgct ggcgcctgct
     7381 gcgcgacggc gagctgctca ccacccactc gccaagctgg aaccgtaaaa aggccgcgtt
     7441 gctggcgttt ttccataggc tccgccgatc ataatcagcc ataccacatt tgtagaggtt
     7501 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     7561 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7621 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7681 atcaatgtat cttatcatgt ctggatcaat tccctatagt gagtcgtatt aaattcgtaa
     7741 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7801 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7861 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgc
//
