LOCUS       sequence-495889-        8846 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for human RECK lacking the CRD, N-terminal 3xFLAG 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8846)
  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 8846)
  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-CRD
FEATURES             Location/Qualifiers
     source          1..8846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1926..2548
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2125..2144)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2572..2592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2590..2888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2942..3736
                     /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(2996..3015)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3606..3625
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4388..4522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4425..4444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4479..4498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4527..4546)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4528..4546)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4560..4576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4560..4576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4573..4595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4584..4600
                     /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(4608..4614,4615..4632,4633..4638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4653..4674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4791..4808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4962..5550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5042..5061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5721..6512,6513..6581))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGRPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6344..6363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6582..6686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6800..6819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7027..7048
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7311..7333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7325..7342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7326..7342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        7503..7882
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7883..8086
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8037..8057
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             8117..8161
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             8162..8227
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 gccaaaatgc ctgcaagaga atcctgatgt ccaagaaaac ggaaatggag attgttgatg
       61 gtctcatcga gggttgtaag acccagccct tgcctcaaga tcctctttgg caatgttttc
      121 ttgaaagctc acaatctgtt caccctggag tcactgtaca ccctcctccc tctacaggcc
      181 tcgatggggc taaattgcat tgttgttcta aagcaaacac ttcaacatgt agggaactct
      241 gcactaaact ttacagcatg agctggggca atacacagag ttggcaagag tttgatcgct
      301 tttgtgaata taatccagtg gaagtgtcca tgttgacctg tttagcggat gtctacctca
      361 ggccaagtac tttaggtaac attgtagaag aagtgactca tccctgtaac ccaaatcctt
      421 gccctgccaa tgagctctgt gaagtaaacc gaaaaggatg tccatctgga gatccctgtc
      481 ttccatactt ttgtgttcaa ggttgcaaac tgggagaagc ttctgatttc attgtccgtc
      541 aagggacact aatccaggtg ccatcatctg caggggaagt tggttgttat aaaatctgtt
      601 catgtggaca aagtggactc ttagaaaact gtatggaaat gcactgtata gacctccaga
      661 agtcttgtat tgttggagga aaaagaaaaa gtcatggaac atcctttagt attgactgca
      721 atgtctgttc ttgttttgct ggcaatttgg tgtgctctac ccgcctttgc ctcagtgagc
      781 acagttcaga agatgaccgt cgtaccttca caggtctgcc ctgtaactgt gcagatcagt
      841 ttgtccctgt atgtgggcag aatgggcgca cttaccccag tgcctgcatt gctcgctgtg
      901 tgggcctcca agaccatcag tttgagtttg gatcatgcat gtcaaaggat ccatgtaatc
      961 ctaatccctg ccaaaaaaac caaaggtgca tacccaaacc acaggtctgc ctgacgactt
     1021 ttgataaatt tggatgtagc cagtatgagt gtgtaccaag acagctcgcg tgtgaccagg
     1081 tccaagatcc tgtttgtgac acagaccaca tggagcacaa caatctctgc actttatacc
     1141 aaagaggaaa aagcctctct tacaaaggtc cctgccagcc cttttgcaga gcaaccgagc
     1201 ccgtatgtgg gcacaatggt gagacctaca gcagtgtgtg tgctgcctac tcggatcgcg
     1261 tggcagtcga ttactatggg gactgccagg ccgtcggagt cctctcagag cacagctccg
     1321 tcgccgagtg tgcttctgtc aagtgtcctt cgctcttggc agctggatgc aaacccatca
     1381 tcccaccggg tgcttgttgc ccattatgtg ctgggatgtt aagagtttta tttgacaaag
     1441 aaaaactgga tactattgct aaggtaacaa ataaaaagcc aataacagtt ctggaaatac
     1501 ttcagaaaat ccgcatgcac gtgtctgtcc cacagtgtga tgtgtttgga tacttcagca
     1561 ttgaatcaga aattgtgatc ctgatcattc ccgtcgatca ctatccaaaa gctctgcaga
     1621 ttgaagcctg caataaagaa gcagagaaga ttgagtccct tatcaactct gacagcccga
     1681 ctttggcgtc ccatgtccct ctctctgccc tcatcatttc ccaggtacag gtctccagca
     1741 gtgtgccatc ggccggtgtc agggccaggc cttcttgcca ctccctcctc cttcccctca
     1801 gcttgggcct tgccttgcac ttgctctgga catataactg actgcccacg gaaagtgcag
     1861 aatgctcctc cacctcactc tcctgccaga tctgatatcg gtaccagtcg actctagagg
     1921 atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     1981 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     2041 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     2101 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     2161 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     2221 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     2281 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     2341 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     2401 ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg
     2461 gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg
     2521 ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg tggaatgtgt
     2581 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2641 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2761 cgcctccgct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2821 ttatttatgc aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2881 ttttttggag gaattgatca gcttgggatc tgatcaagag acaggatgag gatcgtttcg
     2941 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3001 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3061 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3121 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3181 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3241 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3301 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3361 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3421 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3481 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3541 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3601 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3661 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3721 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3781 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3841 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3901 gcccaccccg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag gctggacgac
     3961 ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat
     4021 ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc tttggtcgac
     4081 ccggacggga cgctcctgcg cctgatacag aacgaattgc ttgcaggcat ctcatgagtg
     4141 tgtcttcccg ttttccgcct gaggtcactg cgtggatgga gcgctggcgc ctgctgcgcg
     4201 acggcgagct gctcaccacc cactcgccaa gctggaaccg taaaaaggcc gcgttgctgg
     4261 cgtttttcca taggctccgc cgatcataat cagccatacc acatttgtag aggttttact
     4321 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4381 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4441 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4501 tgtatcttat catgtctgga tcaattccct atagtgagtc gtattaaatt cgtaatcatg
     4561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4621 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcttc cgatcgttgt cagaagtaag
     6181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6601 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6661 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     6721 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6781 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6841 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6901 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6961 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaagag tccactatta
     7021 aagaacgtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     7081 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7141 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag
     7201 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     7261 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     7321 acgttgtaaa acgacggcca gtgccaagct gatctataca ttgaatcaat attggcaatt
     7381 agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac
     7441 gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg
     7501 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     7561 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7621 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7681 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7741 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     7801 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     7861 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     7921 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     7981 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     8041 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     8101 tcagaattaa ttcaccatgt ctgcacttct gatcctagct cttgttggag ctgcagttgc
     8161 tgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
     8221 tgacaagctt gcggccgctg aggtggcagg gggcctggct ccgggcagtg cgggtgcatt
     8281 gtgttgtaat cattcaaagg ataaccaaat gtgccgtgat gtatgtgaac agattttctc
     8341 ctcaaaaagt gaatcccgac taaaacatct gttgcagcga gccccagatt attgcccaga
     8401 gacaatggtt gaaatttgga attgtatgaa ttcatctttg ccaggtgtgt ttaagaagtc
     8461 tgatggctgg gttggcttag gctgctgtga actggctatt gccttggagt gtcgacaggc
     8521 atgcaagcag gcatcttcaa agaatgatat ttccaaagtt tgcagaaaag aatatgagaa
     8581 tgctcttttc agttgcatta gcagaaatga aatgggctcg gtttgttgca gttatgcagg
     8641 tcatcacaca aactgccgag aatactgtca agccattttt cgaacagact cttctcctgg
     8701 tccatctcag ataaaagcag tggaaaatta ttgcgcctct attagtccac aattaataca
     8761 ttgtgtgaac aattatactc aatcttatcc aatgaggaac ccaacggata gtttatattg
     8821 ctgtgacaga gctgaagacc atgctt
//
