LOCUS       sequence-495877-        7533 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted human GPR124 ECD lacking the GAIN 
            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 7533)
  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 7533)
  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-GAIN-His
FEATURES             Location/Qualifiers
     source          1..7533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     476..495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     729..746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    863..884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(899..904,905..922,923..929)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    937..953
                     /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     942..964
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     961..977
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     961..977
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     991..1010
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        991..1009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1015..1149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1039..1058)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1093..1112
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1801..2595)
                     /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(1912..1931)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2522..2541
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2649..2947)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     2945..2965
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2989..3611
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3393..3412
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(3622..3639)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4843..4908)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4909..4953)
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     promoter        complement(4984..5187)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5013..5033)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5188..5567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5728..5745)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5728..5744)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5737..5759)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6022..6043)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6251..6270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6384..6488
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6489..6557,6558..7349)
                     /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(6707..6726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7520..7533,1..575)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
       61 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      121 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      361 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      601 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      661 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      721 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      781 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      841 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      901 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      961 caggaaacag ctatgacatg attacgaatt taatacgact cactataggg aattgatcca
     1021 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     1081 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1141 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1201 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcggcgg
     1261 agcctatgga aaaacgccag caacgcggcc tttttacggt tccagcttgg cgagtgggtg
     1321 gtgagcagct cgccgtcgcg cagcaggcgc cagcgctcca tccacgcagt gacctcaggc
     1381 ggaaaacggg aagacacact catgagatgc ctgcaagcaa ttcgttctgt atcaggcgca
     1441 ggagcgtccc gtccgggtcg accaaagcgg ccatcgtgcc tccccactcc tgcagttcgg
     1501 gggcatggat gcgcggatag ccgctgctgg tttcctggat gccgacggat ttgcactgcc
     1561 ggtagaactc cgcgaggtcg tccagcctca ggcagcagct gaaccaactc gcgaggggat
     1621 cgagcccggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     1681 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     1741 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     1801 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     1861 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     1921 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     1981 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     2041 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     2101 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     2161 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     2221 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     2281 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     2341 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     2401 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     2461 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     2521 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     2581 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ccaagctgat
     2641 caattcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     2701 cggcctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     2761 ggagttagcg gaggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     2821 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     2881 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     2941 ccacacccta actgacacac attccacagc caagctggcc gcgtacccaa ttcaacaggc
     3001 atctactgag tggacccaac gcatgagagg acagtgccaa gcaagcaact caaatgtccc
     3061 accggttggg catggccagg tagcctatgc tgtgtctgga cgtcctcctg ctggtatagt
     3121 tattttaaaa tcagaaggac agggaaggga gcagtggttc acgcctgtaa tcccagcaat
     3181 ttgggaggcc aaggtgggta gatcacctga gattaggagt tggagaccag cctggccaat
     3241 atggtgaaac cccgtctcta ccaaaaaaac aaaaattagc tgagcctggt catgcatgcc
     3301 tggaatccca acaactcggg aggctgaggc aggagaatcg cttgaaccca ggaggcggag
     3361 attgcagtga gccaagattg tgccactgca ctccagcttg gttcccaata gaccccgcag
     3421 gccctacagg ttgtcttccc aacttgcccc ttgctccata ccacccccct ccaccccata
     3481 atattataga aggacaccta gtcagacaaa atgatgcaac ttaattttat taggacaagg
     3541 ctggtgggca ctggagtggc aacttccagg gccaggagag gcactgggga ggggtcacag
     3601 ggatgccacc cgggatcctt aatgatgatg atgatgatga gaacccaccc tggtgatgtc
     3661 gttggtgtag agacagtggg agtagtcccc tggctcccag cggccggcac ggtcacaccg
     3721 gcgggaggct cgggtgcccg gggcaccccc gcccaggggc actgaggtga agggatactg
     3781 caggcaggac tggtaggctg tgatgccagc cagagttcgg ggccacctga agtccccgcg
     3841 gttgttggca acacgctcgg cggggcagta ggaggcagag gtctccagca ccacgatctc
     3901 caccttcttg ctggcgttgc cttgggccat ggacacggtg cactcccact cgcctgaggc
     3961 ccacacgccg atgtgagaca gcgtcagctc actggtgatg aaggtgcagt cgtggatgag
     4021 gctctcggcc aggaggatgc ccgcctgctc atcaccctcc acaggggctc ggttgtggta
     4081 ccagcggatg cgggtgtcgt tgcccaggta gctggcagag cactggaagg gcagccgatc
     4141 cccctggaac accacttggc gtagggacgg gatgaggtgg tgtgtgtgca gctccagggc
     4201 cccctcgcag cagagctggg cctcctggag gctgcccagg gcctgagcat gcagggcact
     4261 ggggtaagca cagagcgtgt gttccgacag ctgcagggag cgattctggg cccagggcag
     4321 cagccagcgc aggtggcagt cacaggtcag gaactcggtg cccaagtcca caaccttaag
     4381 ggctggcagc tcatcaaaga ccccaggttg cagactggag aagatgtttc cagatatgtt
     4441 tagtcggaga agcctgggga ggccctggaa ggtctcggag gtgagacagc caatccggtt
     4501 gttggagaga tctaaacgct tcagctcccc caggcccagg aaggcgcccg gctgcactgt
     4561 gctgatgatg ttgttcctca ggtccagctt ctccagcagt gacagtccca ggaaggagcc
     4621 attgcggagc cccgtgatct tgttattgct caagagcagg gtaacggtgc cgttaggcag
     4681 aaggccgggc tcgggaggct ccgggaggtc cccgccgctg cacaccaccc tccgccgagc
     4741 cgggccaggg acgccgccgc tcagcccctt gggccgctcc cccgagcact tgcagctgcg
     4801 gatggatagc gggcagccgg gcgctgaatt cgcggccgca agcttgtcat cgtcatcctt
     4861 gtaatcgatg tcatgatctt tataatcacc gtcatggtct ttgtagtcag caactgcagc
     4921 tccaacaaga gctaggatca gaagtgcaga catggtgaat taattctgac ggttcactaa
     4981 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaacgg
     5041 ggcggggtta ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     5101 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     5161 attggtgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     5221 ctgccaagta ggaaagtccc gtaaggtcat gtactgggca taatgccagg cgggccattt
     5281 accgtcattg acgtcaatag ggggcggact tggcatatga tacacttgat gtactgccaa
     5341 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     5401 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     5461 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     5521 tgaccccgta attgattact attaataact agtcaataat caatgtcaac atggcggtca
     5581 tattggacat gagccaatat aaatgtacat attatgatat agatacaacg tatgcaatgg
     5641 ccaatagcca atattgattt atgctatata accaatgact aatatggcta attgccaata
     5701 ttgattcaat gtatagatca gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     5761 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5821 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5881 atggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5941 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6001 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cacgttcttt aatagtggac
     6061 tcttaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6121 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6181 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6241 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     6301 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     6361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6481 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6601 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6661 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6721 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6781 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6901 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7021 ctacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7081 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7321 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7501 aaaagatcaa aggatcttct tgagatcctt ttt
//
