LOCUS       sequence-495880-        8597 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted mouse Gpr124 ECD, N-terminal 3xFLAG 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8597)
  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 8597)
  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
FEATURES             Location/Qualifiers
     source          1..8597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1201..1212
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1324..1946
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1523..1542)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1970..1990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1988..2286
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2340..3134
                     /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(2394..2413)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3004..3023
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3786..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3823..3842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3877..3896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3925..3944)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3926..3944)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3958..3974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3958..3974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3971..3993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3982..3998
                     /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(4006..4012,4013..4030,4031..4036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4051..4072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4189..4206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4360..4948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4440..4459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5119..5910,5911..5979))
                     /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     5742..5761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5980..6084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6198..6217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6425..6446
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6709..6731
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6723..6740
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6724..6740
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6901..7280
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7281..7484
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7435..7455
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             7515..7559
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             7560..7625
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cgggtgacca acaaccgtgg ggacttcagg tggccccgaa ccttggctgg catcacggct
       61 taccagtcct gtttacagta ccccttcacc tctgtgccct tgagtggggg agccccgggt
      121 acccgagcct cacgcaggtg tgaccgagct ggccgctggg agccagggga ctactcgcac
      181 tgtctgtaca ccaatgacat cactcgggtg ctctacacct ttgtgctgat gcccatcaac
      241 gcctccaatg cattgacgtt ggcccaccag ctgcgagtgt ataccgcaga ggccgccagc
      301 ttctcagaca tgatggacgt ggtctatgtg gctcagatga tccagaagtt tttgggttac
      361 gttgaccaga tcaaggagct ggtggaggtg atggtggaca tggccagcaa cctgatgctg
      421 gtggatgagc accttctgtg gctggcccag agagaagaca aagcctgcag tggcattgtg
      481 ggtgccctgg agcgaatcgg aggagctgct cttagccccc atgcccagca catctctgtg
      541 aattcaagaa atgtggcact ggaggcctac ctcatcaagc ctcacagcta cgtgggtctg
      601 acttgcacgg ctttccagag gagggaggta ggagtgtcgg gtgcacagcc aagcagcgtc
      661 ggccaggatg ccccagtgga gcccgaaccc ctagctgatc agcagcttag gttccgctgc
      721 accactggga ggcccaacat ttctctgtca tccttccaca tcaagaatag cgtggccctg
      781 gcctccatcc agctgccccc cagcctgttc tcaacccttc cggctgccct ggctccccca
      841 gtccctccag attgcaccct gcaactgctg gtcttcagaa atggccgtct tttccgcagc
      901 cacggcaaca acacttcccg tcctggagca gctgggcctg gcaagaggcg tggtgtggcc
      961 accccagtca tatttgcagg aaccagtggc tgtggtgtgg gaaacttgac ggagcccgtg
     1021 gctgtgtcac tgaggcactg ggctgaagga gctgacccca tggcagcttg gtggaaccag
     1081 gacgggcctg gaggctggag ttctgaaggc tgcaggctcc gctacagcca gcccaacgtc
     1141 agctccctgt actgccagca cttgggcaat gtggccgtgc ttatggagct gtaacggccg
     1201 atagaagggc gaattccagc acactggcgg ccgttactag tggatccgag ctcggtacca
     1261 agcttgcggc cgcgaattca tcgatagatc tgatatcggt accagtcgac tctagaggat
     1321 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     1381 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     1441 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     1501 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     1561 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     1621 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     1681 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     1741 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     1801 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc
     1861 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg
     1921 tccactcagt agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt
     1981 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2041 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2101 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2161 cctccgctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2221 atttatgcag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2281 ttttggagga attgatcagc ttgggatctg atcaagagac aggatgagga tcgtttcgca
     2341 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2401 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2461 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2521 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2581 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2641 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2701 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2761 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     2821 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     2881 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     2941 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3001 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3061 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3121 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3181 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3241 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3301 ccaccccggg ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc tggacgacct
     3361 cgcggagttc taccggcagt gcaaatccgt cggcatccag gaaaccagca gcggctatcc
     3421 gcgcatccat gcccccgaac tgcaggagtg gggaggcacg atggccgctt tggtcgaccc
     3481 ggacgggacg ctcctgcgcc tgatacagaa cgaattgctt gcaggcatct catgagtgtg
     3541 tcttcccgtt ttccgcctga ggtcactgcg tggatggagc gctggcgcct gctgcgcgac
     3601 ggcgagctgc tcaccaccca ctcgccaagc tggaaccgta aaaaggccgc gttgctggcg
     3661 tttttccata ggctccgccg atcataatca gccataccac atttgtagag gttttacttg
     3721 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3781 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3841 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3901 tatcttatca tgtctggatc aattccctat agtgagtcgt attaaattcg taatcatgtc
     3961 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4021 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4801 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5401 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5701 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     6121 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6181 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6241 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6301 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6361 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaagagtc cactattaaa
     6421 gaacgtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6481 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6541 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc
     6601 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6661 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6721 gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag
     6781 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     6841 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
     6901 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6961 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     7021 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     7081 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     7141 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     7201 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
     7261 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
     7321 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     7381 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
     7441 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     7501 agaattaatt caccatgtct gcacttctga tcctagctct tgttggagct gcagttgctg
     7561 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg
     7621 acaagcttgc gcctggctgc ccggtcccta tccgcggttg caagtgctct ggggagcggc
     7681 ccaagggact aagtggcggc gcccacaacc cggctcgaag gagggtggtg tgcggcagtg
     7741 gggatctccc cgaacctcca gatcccggcc ttctgccaaa cggcaccatc accttgctct
     7801 tgagcaacaa caagattact gggctccgca atggatcctt cttgggactg tccctgttgg
     7861 agaagttgga cctgaggagc aatgtcatca gcactgtgca gcctggagcc ttcctaggtc
     7921 tgggagagct aaaacgctta gatctctcca acaatcggat tggctgtctc acctctgaga
     7981 catttcaagg gctccctaga cttctcagac taaacatatc tggaaacatc tactctagtc
     8041 tgcaacctgg ggtctttgat gagctgccag cccttaagat tgtggacttt ggtactgagt
     8101 ttctgacctg tgactgccgc ctgcgctggc tgctgccctg ggcccggaat cactccctgc
     8161 agctgtctga gcgcacactc tgtgcctacc ccagtgccct gcacgcccat gccctgagca
     8221 gcctccagga gtcccagctt cgctgtgaag gggccctgga actgcacacc cactacctca
     8281 tcccatccct tcgccaagtg gtgttccagg gtgaccgcct gcccttccag tgctcagcca
     8341 gctacttggg caacgatacc cggatccact ggtaccacaa tggggctcct atggagagcg
     8401 atgagcaggc cggcatcgtc cttgctgaaa accttatcca tgattgcacc ttcatcacca
     8461 gtgagctgac cctgtctcac attggtgtgt gggcctctgg tgaatgggag tgctccgtgt
     8521 ccacagtcca aggcaacacc agcaagaagg tggagatagt agtgctggag acctctgcct
     8581 cctactgccc tgcagag
//
