LOCUS       sequence-495890-        7994 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted human RECK lacking CC domains 1-5, 
            fused to mouse IgG2a Fc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7994)
  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 7994)
  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-sRECK Delta-CK-Fc
FEATURES             Location/Qualifiers
     source          1..7994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(147..166)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        338..717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        718..921
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     872..892
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     966..985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        966..984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1008..1067
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2952..3647
                     /codon_start=1
                     /product="CH2 and CH3 domains of the mouse IgG2a heavy 
                     chain plus the hinge region"
                     /label=Fc tag
                     /translation="PRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVT
                     CVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEF
                     KCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIY
                     VEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHT
                     TKSFSRTPGK"
     primer_bind     complement(3688..3705)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3694..3918
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3964..4392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4051..4070)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4261..4282
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4401..4421)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4406..4735
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4586..4721
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4648..4667
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4802..5596
                     /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(4856..4875)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5466..5485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5770..5891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5807..5826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5861..5880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5940..5956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5940..5956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5953..5975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5964..5980
                     /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(5988..5994,5995..6012,6013..6018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6033..6054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6171..6188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6342..6930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6422..6441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7101..7892,7893..7961))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASWQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7724..7743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7962..7994,1..72))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
       61 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
      121 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
      181 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
      241 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
      301 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      361 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      421 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      481 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      541 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      601 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      661 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      721 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      781 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      841 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      901 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
      961 gaaattaata cgactcacta tagggagacc caagctggct agccaccatg gagacagaca
     1021 cactcctgct atgggtactg ctgctctggg ttccaggttc cactggtgac gcggcccagc
     1081 cggccgaggt ggcagggggc ctggctccgg gcagtgcggg tgcattgtta gcggatgtcc
     1141 gggaaccttg ccagttgggc tgtagaaacc ttacttactg tactaatttt aacaacaggc
     1201 caacagaact tttcaggagt tgtaatgcac agtcagatca aggagccatg aatgacatga
     1261 agttgtggga gaaaggaagc ataaagatgc catttatcaa tatacctgtt cttgatatta
     1321 aaaagtgcca gccagagatg tggaaagcaa tagcttgttc actgcagatt aaaccttgtc
     1381 atagtaaatc tcggggaagt attatttgca aatcagattg tgtggagatt cttaaaaaat
     1441 gtggagacca gaacaaattc cctgaagacc acacagctga aagtatttgt gagcttctgt
     1501 cacctacaga tgatctgaag aattgtatac ctttggatac atacctcagg ccaagtactt
     1561 taggtaacat tgtagaagaa gtgactcatc cctgtaaccc aaatccttgc cctgccaatg
     1621 agctctgtga agtaaaccga aaaggatgtc catctggaga tccctgtctt ccatactttt
     1681 gtgttcaagg ttgcaaactg ggagaagctt ctgatttcat tgtccgtcaa gggacactaa
     1741 tccaggtgcc atcatctgca ggggaagttg gttgttataa aatctgttca tgtggacaaa
     1801 gtggactctt agaaaactgt atggaaatgc actgtataga cctccagaag tcttgtattg
     1861 ttggaggaaa aagaaaaagt catggaacat cctttagtat tgactgcaat gtctgttctt
     1921 gttttgctgg caatttggtg tgctctaccc gcctttgcct cagtgagcac agttcagaag
     1981 atgaccgtcg taccttcaca ggtctgccct gtaactgtgc agatcagttt gtccctgtat
     2041 gtgggcagaa tgggcgcact taccccagtg cctgcattgc tcgctgtgtg ggcctccaag
     2101 accatcagtt tgagtttgga tcatgcatgt caaaggatcc atgtaatcct aatccctgcc
     2161 aaaaaaacca aaggtgcata cccaaaccac aggtctgcct gacgactttt gataaatttg
     2221 gatgtagcca gtatgagtgt gtaccaagac agctcgcgtg tgaccaggtc caagatcctg
     2281 tttgtgacac agaccacatg gagcacaaca atctctgcac tttataccaa agaggaaaaa
     2341 gcctctctta caaaggtccc tgccagccct tttgcagagc aaccgagccc gtatgtgggc
     2401 acaatggtga gacctacagc agtgtgtgtg ctgcctactc ggatcgcgtg gcagtcgatt
     2461 actatgggga ctgccaggcc gtcggagtcc tctcagagca cagctccgtc gccgagtgtg
     2521 cttctgtcaa gtgtccttcg ctcttggcag ctggatgcaa acccatcatc ccaccgggtg
     2581 cttgttgccc attatgtgct gggatgttaa gagttttatt tgacaaagaa aaactggata
     2641 ctattgctaa ggtaacaaat aaaaagccaa taacagttct ggaaatactt cagaaaatcc
     2701 gcatgcacgt gtctgtccca cagtgtgatg tgtttggata cttcagcatt gaatcagaaa
     2761 ttgtgatcct gatcattccc gtcgatcact atccaaaagc tctgcagatt gaagcctgca
     2821 ataaagaagc agagaagatt gagtccctta tcaactctga cagcccgact ttggcgtccc
     2881 atgtccctct ctctgccctc atcatttccc aggtacaggt ctccagcagt gtgccatcga
     2941 ctcgactaga gcccagaggg cccacaatca agccctgtcc tccatgcaaa tgcccagcac
     3001 ctaacctctt gggtggacca tccgtcttca tcttccctcc aaagatcaag gatgtactca
     3061 tgatctccct gagccccata gtcacatgtg tggtggtgga tgtgagcgag gatgacccag
     3121 atgtccagat cagctggttt gtgaacaacg tggaagtaca cacagctcag acacaaaccc
     3181 atagagagga ttacaacagt actctccggg tggtcagtgc cctccccatc cagcaccagg
     3241 actggatgag tggcaaggag ttcaaatgca aggtcaacaa caaagacctc ccagcgccca
     3301 tcgagagaac catctcaaaa cccaaagggt cagtaagagc tccacaggta tatgtcttgc
     3361 ctccaccaga agaagagatg actaagaaac aggtcactct gacctgcatg gtcacagact
     3421 tcatgcctga agacatttac gtggagtgga ccaacaacgg gaaaacagag ctaaactaca
     3481 agaacactga accagtcctg gactctgatg gttcttactt catgtacagc aagctgagag
     3541 tggaaaagaa gaactgggtg gaaagaaata gctactcctg ttcagtggtc cacgagggtc
     3601 tgcacaatca ccacacgact aagagcttct cccggactcc gggtaaatga ctcgagtcta
     3661 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3721 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3781 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3841 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3901 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3961 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4021 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4081 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4141 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4201 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4261 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4321 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4381 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4441 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4501 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4561 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4621 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4681 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4741 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4801 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4861 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4921 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4981 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5041 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5101 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5161 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5221 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5281 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5341 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5401 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5461 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5521 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5581 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5641 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5701 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5761 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5821 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5881 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5941 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6001 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6061 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6121 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6181 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6241 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7321 actttatccg cctccatcca gtctattaat tgttgccagg aagctagagt aagtagttcg
     7381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7981 tattgaagca ttta
//
