LOCUS       sequence-495885-        8342 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for human RECK lacking CC domains 1-5, N-terminal 
            3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8342)
  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 8342)
  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-RECK Delta-CK
FEATURES             Location/Qualifiers
     source          1..8342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1176..1798
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1375..1394)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1822..1842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1840..2138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2192..2986
                     /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(2246..2265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2856..2875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3638..3772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3675..3694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3729..3748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3777..3796)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3778..3796)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3810..3826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3810..3826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3823..3845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3834..3850
                     /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(3858..3864,3865..3882,3883..3888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3903..3924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4041..4058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4212..4800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4292..4311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4971..5762,5763..5831))
                     /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     5594..5613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5832..5936)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6050..6069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6277..6298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6561..6583
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6575..6592
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6576..6592
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6753..7132
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7133..7336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7287..7307
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             7367..7411
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             7412..7477
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 tgtgctctac ccgcctttgc ctcagtgagc acagttcaga agatgaccgt cgtaccttca
       61 caggtctgcc ctgtaactgt gcagatcagt ttgtccctgt atgtgggcag aatgggcgca
      121 cttaccccag tgcctgcatt gctcgctgtg tgggcctcca agaccatcag tttgagtttg
      181 gatcatgcat gtcaaaggat ccatgtaatc ctaatccctg ccaaaaaaac caaaggtgca
      241 tacccaaacc acaggtctgc ctgacgactt ttgataaatt tggatgtagc cagtatgagt
      301 gtgtaccaag acagctcgcg tgtgaccagg tccaagatcc tgtttgtgac acagaccaca
      361 tggagcacaa caatctctgc actttatacc aaagaggaaa aagcctctct tacaaaggtc
      421 cctgccagcc cttttgcaga gcaaccgagc ccgtatgtgg gcacaatggt gagacctaca
      481 gcagtgtgtg tgctgcctac tcggatcgcg tggcagtcga ttactatggg gactgccagg
      541 ccgtcggagt cctctcagag cacagctccg tcgccgagtg tgcttctgtc aagtgtcctt
      601 cgctcttggc agctggatgc aaacccatca tcccaccggg tgcttgttgc ccattatgtg
      661 ctgggatgtt aagagtttta tttgacaaag aaaaactgga tactattgct aaggtaacaa
      721 ataaaaagcc aataacagtt ctggaaatac ttcagaaaat ccgcatgcac gtgtctgtcc
      781 cacagtgtga tgtgtttgga tacttcagca ttgaatcaga aattgtgatc ctgatcattc
      841 ccgtcgatca ctatccaaaa gctctgcaga ttgaagcctg caataaagaa gcagagaaga
      901 ttgagtccct tatcaactct gacagcccga ctttggcgtc ccatgtccct ctctctgccc
      961 tcatcatttc ccaggtacag gtctccagca gtgtgccatc ggccggtgtc agggccaggc
     1021 cttcttgcca ctccctcctc cttcccctca gcttgggcct tgccttgcac ttgctctgga
     1081 catataactg actgcccacg gaaagtgcag aatgctcctc cacctcactc tcctgccaga
     1141 tctgatatcg gtaccagtcg actctagagg atcccgggtg gcatccctgt gacccctccc
     1201 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     1261 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     1321 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     1381 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     1441 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     1501 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     1561 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     1621 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     1681 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     1741 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     1801 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1981 ccatagtccc gcccctaact ccgcccatcc cgcctccgct aactccgccc agttccgccc
     2041 attctccgcc ccatggctga ctaatttttt ttatttatgc aggccgaggc cgcctcggcc
     2101 tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca gcttgggatc
     2161 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2221 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2281 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2341 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2401 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2461 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2521 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2581 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2641 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2701 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2761 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2821 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2881 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2941 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3001 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3061 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3121 cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt
     3181 tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca gtgcaaatcc
     3241 gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga actgcaggag
     3301 tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg cctgatacag
     3361 aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg
     3421 cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc cactcgccaa
     3481 gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc cgatcataat
     3541 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3601 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaattccct
     3781 atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     3841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4021 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4081 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4621 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4681 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4741 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4861 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4921 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4981 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5041 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5101 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5161 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5221 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5281 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5341 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5401 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5461 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5521 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5581 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5641 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5701 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5761 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5821 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5941 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6001 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6061 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6121 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6181 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6241 gagtccacgt tctttaagag tccactatta aagaacgtgg actcttgttc caaactggaa
     6301 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6361 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6421 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6481 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6541 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     6601 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
     6661 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
     6721 tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata
     6781 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6841 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6901 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6961 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
     7021 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
     7081 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     7141 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     7201 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     7261 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
     7321 ctatataagc agagctcgtt tagtgaaccg tcagaattaa ttcaccatgt ctgcacttct
     7381 gatcctagct cttgttggag ctgcagttgc tgactacaaa gaccatgacg gtgattataa
     7441 agatcatgac atcgattaca aggatgacga tgacaagctt gcggccgctg aggtggcagg
     7501 gggcctggct ccgggcagtg cgggtgcatt gttagcggat gtccgggaac cttgccagtt
     7561 gggctgtaga aaccttactt actgtactaa ttttaacaac aggccaacag aacttttcag
     7621 gagttgtaat gcacagtcag atcaaggagc catgaatgac atgaagttgt gggagaaagg
     7681 aagcataaag atgccattta tcaatatacc tgttcttgat attaaaaagt gccagccaga
     7741 gatgtggaaa gcaatagctt gttcactgca gattaaacct tgtcatagta aatctcgggg
     7801 aagtattatt tgcaaatcag attgtgtgga gattcttaaa aaatgtggag accagaacaa
     7861 attccctgaa gaccacacag ctgaaagtat ttgtgagctt ctgtcaccta cagatgatct
     7921 gaagaattgt atacctttgg atacatacct caggccaagt actttaggta acattgtaga
     7981 agaagtgact catccctgta acccaaatcc ttgccctgcc aatgagctct gtgaagtaaa
     8041 ccgaaaagga tgtccatctg gagatccctg tcttccatac ttttgtgttc aaggttgcaa
     8101 actgggagaa gcttctgatt tcattgtccg tcaagggaca ctaatccagg tgccatcatc
     8161 tgcaggggaa gttggttgtt ataaaatctg ttcatgtgga caaagtggac tcttagaaaa
     8221 ctgtatggaa atgcactgta tagacctcca gaagtcttgt attgttggag gaaaaagaaa
     8281 aagtcatgga acatccttta gtattgactg caatgtctgt tcttgttttg ctggcaattt
     8341 gg
//
