LOCUS       sequence-495878-        6996 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted human GPR124 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 6996)
  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 6996)
  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 LRR-His
FEATURES             Location/Qualifiers
     source          1..6996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             481..498
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    509..1131
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(708..727)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1155..1175)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1173..1471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1525..2319
                     /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(1579..1598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2189..2208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2971..3105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3008..3027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3062..3081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3110..3129)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3111..3129)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3143..3159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3143..3159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3156..3178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3167..3183
                     /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(3191..3197,3198..3215,3216..3221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3236..3257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3374..3391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3545..4133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3625..3644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4304..5095,5096..5164))
                     /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     4927..4946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5165..5269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5383..5402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5610..5631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5894..5916
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5908..5925
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5909..5925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6086..6465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6466..6669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6620..6640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             6700..6744
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             6745..6810
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 ctcttgagca ataacaagat cacggggctc cgcaatggct ccttcctggg actgtcactg
       61 ctggagaagc tggacctgag gaacaacatc atcagcacag tgcagccggg cgccttcctg
      121 ggcctggggg agctgaagcg tttagatctc tccaacaacc ggattggctg tctcacctcc
      181 gagaccttcc agggcctccc caggcttctc cgactaaaca tatctggaaa catcttctcc
      241 agtctgcaac ctggggtctt tgatgagctg ccagccctta aggttgtgga cttgggcacc
      301 gagttcctga cctgtgactg ccacctgcgc tggctgctgc cctgggccca gaatcgctcc
      361 ctgcagctgt cggaacacac gctctgtgct taccccagtg ccctgcatgc tcaggccctg
      421 ggcagcctcc aggaggccca gctctgctgc gagggggccc tggagctgca cacaggttct
      481 catcatcatc atcatcatta aggatcccgg gtggcatccc tgtgacccct ccccagtgcc
      541 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
      601 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
      661 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
      721 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
      781 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
      841 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
      901 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
      961 aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt
     1021 ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct
     1081 tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg
     1141 gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1201 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1261 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     1321 cccgccccta actccgccca tcccgcctcc gctaactccg cccagttccg cccattctcc
     1381 gccccatggc tgactaattt tttttattta tgcaggccga ggccgcctcg gcctctgagc
     1441 tattccagaa gtagtgagga ggcttttttg gaggaattga tcagcttggg atctgatcaa
     1501 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     1561 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     1621 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     1681 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     1741 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     1801 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     1861 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     1921 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     1981 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2041 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2101 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2161 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2221 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     2281 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     2341 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     2401 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     2461 gggatctcat gctggagttc ttcgcccacc ccgggctcga tcccctcgcg agttggttca
     2521 gctgctgcct gaggctggac gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca
     2581 tccaggaaac cagcagcggc tatccgcgca tccatgcccc cgaactgcag gagtggggag
     2641 gcacgatggc cgctttggtc gacccggacg ggacgctcct gcgcctgata cagaacgaat
     2701 tgcttgcagg catctcatga gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat
     2761 ggagcgctgg cgcctgctgc gcgacggcga gctgctcacc acccactcgc caagctggaa
     2821 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgccgatcat aatcagccat
     2881 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2941 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3061 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaattc cctatagtga
     3121 gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3181 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3241 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3301 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4381 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4501 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4741 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4801 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4921 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4981 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5041 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5101 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5161 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5221 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5281 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5581 cgttctttaa gagtccacta ttaaagaacg tggactcttg ttccaaactg gaacaacact
     5641 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5701 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5761 tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5821 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5881 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctgatctat
     5941 acattgaatc aatattggca attagccata ttagtcattg gttatatagc ataaatcaat
     6001 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     6061 tcatgtccaa tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca
     6121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     6181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     6241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
     6361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
     6421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     6481 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     6541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6601 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6661 agcagagctc gtttagtgaa ccgtcagaat taattcacca tgtctgcact tctgatccta
     6721 gctcttgttg gagctgcagt tgctgactac aaagaccatg acggtgatta taaagatcat
     6781 gacatcgatt acaaggatga cgatgacaag cttgcggccg cgaattcagc gcccggctgc
     6841 ccgctatcca tccgcagctg caagtgctcg ggggagcggc ccaaggggct gagcggcggc
     6901 gtccctggcc cggctcggcg gagggtggtg tgcagcggcg gggacctccc ggagcctccc
     6961 gagcccggcc ttctgcctaa cggcaccgtt accctg
//
