LOCUS       sequence-495859-        8325 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for a secreted human GPR124 ECD-mouse IgG2a Fc 
            fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8325)
  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 8325)
  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: pcDNA3.1-Ig-kappa SP-sGPR124-Fc
FEATURES             Location/Qualifiers
     source          1..8325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1955..1972)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1961..2185
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2231..2659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2318..2337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2528..2549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2668..2688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2673..3002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2853..2988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2915..2934
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3069..3863
                     /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(3123..3142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3733..3752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4037..4158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4074..4093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4128..4147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4207..4223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4207..4223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4220..4242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4231..4247
                     /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(4255..4261,4262..4279,4280..4285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4300..4321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4438..4455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4609..5197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4689..4708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5368..6159,6160..6228))
                     /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     5991..6010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6229..6333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6408..6427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6599..6978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6979..7182
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7133..7153
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7227..7246
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7227..7245
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7287..7296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agcgtgttgc caacaaccgc ggggacttca ggtggccccg aactctggct ggcatcacag
       61 cctaccagtc ctgcctgcag tatcccttca cctcagtgcc cctgggcggg ggtgccccgg
      121 gcacccgagc ctcccgccgg tgtgaccgtg ccggccgctg ggagccaggg gactactccc
      181 actgtctcta caccaacgac atcaccaggg tgctgtacac cttcgtgctg atgcccatca
      241 atgcctccaa tgcgctgacc ctggctcacc agctgcgcgt gtacacagcc gaggccgcta
      301 gcttttcaga catgatggat gtagtctatg tggctcagat gatccagaaa tttttgggtt
      361 atgtcgacca gatcaaagag ctggtagagg tgatggtgga catggccagc aacctgatgc
      421 tggtggacga gcacctgctg tggctggccc agcgcgagga caaggcctgc agccgcatcg
      481 tgggtgccct ggagcgcatt gggggggccg ccctcagccc ccatgcccag cacatctcag
      541 tgaatgcgag gaacgtggca ttggaggcct acctcatcaa gccgcacagc tacgtgggcc
      601 tgacctgcac agccttccag aggagggagg gaggggtgcc gggcacacgg ccaggaagcc
      661 ctggccagaa ccccccacct gagcccgagc ccccagctga ccagcagctc cgcttccgct
      721 gcaccaccgg gaggcccaat gtttctctgt cgtccttcca catcaagaac agcgtggccc
      781 tggcctccat ccagctgccc ccgagtctat tctcatccct tccggctgcc ctggctcccc
      841 cggtgccccc agactgcacc ctgcaactgc tcgtcttccg aaatggccgc ctcttccaca
      901 gccacagcaa cacctcccgc cctggagctg ctgggcctgg caagaggcgt ggcgtggcca
      961 cccccgtcat cttcgcagga accagtggct gtggcgtggg aaacctgaca gagccagtgg
     1021 ccgtttcgct gcggcactgg gctgagggag ccgaacctgt ggccgcttgg tggagccagg
     1081 aggggcccgg ggaggctggg ggctggacct cggagggctg ccagctccgc tccagccagc
     1141 ccaatgtcag cgccctgcac tgccagcact tgggcaatgt ggccgtgctc atggagctga
     1201 ctcgactaga gcctagaggc cctaccatca agccctgtcc tccatgcaaa tgccccgctc
     1261 ctaatctgct cggcggaccc agcgtgttca tcttcccacc taagatcaag gacgtgctga
     1321 tgatctctct gagccccatc gtgacctgcg tggtggtgga tgtgtccgag gacgatcccg
     1381 atgtgcagat cagttggttc gtgaacaacg tggaagtgca cacagcccag acacagaccc
     1441 acagagagga ctacaacagc accctgagag tggtgtctgc cctgcctatc cagcaccagg
     1501 attggatgag cggcaaagaa ttcaagtgca aagtgaacaa caaggacctg cctgctccta
     1561 tcgagcggac catctctaag cctaagggct ctgttagagc ccctcaggtg tacgtgctgc
     1621 ctcctccaga ggaagagatg accaagaaac aagtgaccct gacctgcatg gtcaccgact
     1681 tcatgcccga ggacatctac gtggaatgga ccaacaacgg caagaccgag ctgaactaca
     1741 agaacaccga gcctgtgctg gacagcgacg gcagctactt catgtacagc aagctgcgcg
     1801 tggaaaagaa gaactgggtc gagcggaaca gctacagctg ctctgtggtg cacgagggcc
     1861 tgcacaatca ccacaccacc aagagcttca gcagaacccc tggcaagtga gcggccgctc
     1921 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     1981 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2041 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2101 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2161 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2221 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2281 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2581 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2641 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2821 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2881 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2941 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3001 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3061 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3121 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3181 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3241 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3301 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3361 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3421 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3481 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3541 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3601 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3661 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3721 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3781 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3841 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3901 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3961 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4021 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4081 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4141 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4201 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4261 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4321 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6361 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     6421 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     6481 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     6541 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     6601 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     6661 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     6721 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     6781 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     6841 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     6901 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     6961 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     7021 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     7081 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     7141 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
     7201 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
     7261 tagcgtttaa acttaagctt ggtaccgcca ccatggaaac cgacacactg ctgctgtggg
     7321 tgctgcttct ttgggtgccc ggatctacag gcgccgctgc gcccggctgc ccgctatcca
     7381 tccgcagctg caagtgctcg ggggagcggc ccaaggggct gagcggcggc gtccctggcc
     7441 cggctcggcg gagggtggtg tgcagcggcg gggacctacc ggagcctccc gagcccggcc
     7501 ttctgcctaa cggcaccgtt accctgctct tgagcaataa caagatcacg gggctccgca
     7561 atggctcctt cctgggactg tcactgctgg agaagctgga cctgaggaac aacatcatca
     7621 gcacagtgca gccgggcgcc ttcctgggcc tgggggagct gaagcgttta gatctctcca
     7681 acaaccggat tggctgtctc acctccgaga ccttccaggg cctccccagg cttctccgac
     7741 taaacatatc tggaaacatc ttctccagtc tgcaacctgg ggtctttgat gagctgccag
     7801 cccttaaggt tgtggacttg ggcaccgagt tcctgacctg tgactgccac ctgcgctggc
     7861 tgctgccctg ggcccagaat cgctccctgc agctgtcgga acacacgctc tgtgcttacc
     7921 ccagtgccct gcatgctcag gccctgggca gcctccagga ggcccagctc tgctgcgagg
     7981 gggccctgga gctgcacaca caccacctca tcccgtccct acgccaagtg gtgttccagg
     8041 gggatcggct gcccttccag tgctctgcca gctacctggg caacgacacc cgcatccgct
     8101 ggtaccacaa ccgagcccct gtggagggtg atgagcaggc gggcatcctc ctggccgaga
     8161 gcctcatcca cgactgcacc ttcatcacca gtgagctgac gctgtctcac atcggcgtgt
     8221 gggcctcagg cgagtgggag tgcaccgtgt ccatggccca aggcaacgcc agcaagaagg
     8281 tggagatcgt ggtgctggag acctctgcct cctactgccc cgccg
//
