LOCUS       sequence-495891-        8498 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for secreted human RECK lacking the CRD, fused to 
            mouse IgG2a Fc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8498)
  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 8498)
  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-CRD-Fc
FEATURES             Location/Qualifiers
     source          1..8498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..26
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    143..164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(179..184,185..202,203..209)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    217..233
                     /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)."
     primer_bind     222..244
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     241..257
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     241..257
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    306..427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(317..336)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     371..390
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(601..1395)
                     /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(712..731)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1322..1341
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1462..1791)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1476..1611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1530..1549)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1776..1796
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1805..2233)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1915..1936)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2127..2146
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2279..2503
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2492..2509
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2550..3245)
                     /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"
     sig_peptide     complement(5634..5693)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      5690..5699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5716..5735)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5717..5735)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5780..5983)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5809..5829)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5984..6363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6535..6554
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6629..6733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6734..6802,6803..7594)
                     /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     complement(6952..6971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7765..8353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8254..8273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
       61 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      121 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      181 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      241 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
      301 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      361 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      421 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
      481 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
      541 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
      601 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
      661 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
      721 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
      781 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
      841 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
      901 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
      961 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     1021 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     1081 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     1141 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     1201 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     1261 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     1321 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     1381 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc cgaaaatgga
     1441 tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     1501 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
     1561 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     1621 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     1681 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     1741 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     1801 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1981 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2041 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2101 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2161 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2221 ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc ctcagaagcc
     2281 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
     2341 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     2401 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     2461 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
     2521 gtttaaacgg gccctctaga ctcgagtcat ttacccggag tccgggagaa gctcttagtc
     2581 gtgtggtgat tgtgcagacc ctcgtggacc actgaacagg agtagctatt tctttccacc
     2641 cagttcttct tttccactct cagcttgctg tacatgaagt aagaaccatc agagtccagg
     2701 actggttcag tgttcttgta gtttagctct gttttcccgt tgttggtcca ctccacgtaa
     2761 atgtcttcag gcatgaagtc tgtgaccatg caggtcagag tgacctgttt cttagtcatc
     2821 tcttcttctg gtggaggcaa gacatatacc tgtggagctc ttactgaccc tttgggtttt
     2881 gagatggttc tctcgatggg cgctgggagg tctttgttgt tgaccttgca tttgaactcc
     2941 ttgccactca tccagtcctg gtgctggatg gggagggcac tgaccacccg gagagtactg
     3001 ttgtaatcct ctctatgggt ttgtgtctga gctgtgtgta cttccacgtt gttcacaaac
     3061 cagctgatct ggacatctgg gtcatcctcg ctcacatcca ccaccacaca tgtgactatg
     3121 gggctcaggg agatcatgag tacatccttg atctttggag ggaagatgaa gacggatggt
     3181 ccacccaaga ggttaggtgc tgggcatttg catggaggac agggcttgat tgtgggccct
     3241 ctgggctcta gtcgagtcga tggcacactg ctggagacct gtacctggga aatgatgagg
     3301 gcagagagag ggacatggga cgccaaagtc gggctgtcag agttgataag ggactcaatc
     3361 ttctctgctt ctttattgca ggcttcaatc tgcagagctt ttggatagtg atcgacggga
     3421 atgatcagga tcacaatttc tgattcaatg ctgaagtatc caaacacatc acactgtggg
     3481 acagacacgt gcatgcggat tttctgaagt atttccagaa ctgttattgg ctttttattt
     3541 gttaccttag caatagtatc cagtttttct ttgtcaaata aaactcttaa catcccagca
     3601 cataatgggc aacaagcacc cggtgggatg atgggtttgc atccagctgc caagagcgaa
     3661 ggacacttga cagaagcaca ctcggcgacg gagctgtgct ctgagaggac tccgacggcc
     3721 tggcagtccc catagtaatc gactgccacg cgatccgagt aggcagcaca cacactgctg
     3781 taggtctcac cattgtgccc acatacgggc tcggttgctc tgcaaaaggg ctggcaggga
     3841 cctttgtaag agaggctttt tcctctttgg tataaagtgc agagattgtt gtgctccatg
     3901 tggtctgtgt cacaaacagg atcttggacc tggtcacacg cgagctgtct tggtacacac
     3961 tcatactggc tacatccaaa tttatcaaaa gtcgtcaggc agacctgtgg tttgggtatg
     4021 cacctttggt ttttttggca gggattagga ttacatggat cctttgacat gcatgatcca
     4081 aactcaaact gatggtcttg gaggcccaca cagcgagcaa tgcaggcact ggggtaagtg
     4141 cgcccattct gcccacatac agggacaaac tgatctgcac agttacaggg cagacctgtg
     4201 aaggtacgac ggtcatcttc tgaactgtgc tcactgaggc aaaggcgggt agagcacacc
     4261 aaattgccag caaaacaaga acagacattg cagtcaatac taaaggatgt tccatgactt
     4321 tttctttttc ctccaacaat acaagacttc tggaggtcta tacagtgcat ttccatacag
     4381 ttttctaaga gtccactttg tccacatgaa cagattttat aacaaccaac ttcccctgca
     4441 gatgatggca cctggattag tgtcccttga cggacaatga aatcagaagc ttctcccagt
     4501 ttgcaacctt gaacacaaaa gtatggaaga cagggatctc cagatggaca tccttttcgg
     4561 tttacttcac agagctcatt ggcagggcaa ggatttgggt tacagggatg agtcacttct
     4621 tctacaatgt tacctaaagt acttggcctg aggtagacat ccgctaaaca ggtcaacatg
     4681 gacacttcca ctggattata ttcacaaaag cgatcaaact cttgccaact ctgtgtattg
     4741 ccccagctca tgctgtaaag tttagtgcag agttccctac atgttgaagt gtttgcttta
     4801 gaacaacaat gcaatttagc cccatcgagg cctgtagagg gaggagggtg tacagtgact
     4861 ccagggtgaa cagattgtga gctttcaaga aaacattgcc aaagaggatc ttgaggcaag
     4921 ggctgggtct tacaaccctc gatgagacca tcaacaatct ccatttccgt tttcttggac
     4981 atcaggattc tcttgcaggc attttggcaa gcatggtctt cagctctgtc acagcaatat
     5041 aaactatccg ttgggttcct cattggataa gattgagtat aattgttcac acaatgtatt
     5101 aattgtggac taatagaggc gcaataattt tccactgctt ttatctgaga tggaccagga
     5161 gaagagtctg ttcgaaaaat ggcttgacag tattctcggc agtttgtgtg atgacctgca
     5221 taactgcaac aaaccgagcc catttcattt ctgctaatgc aactgaaaag agcattctca
     5281 tattcttttc tgcaaacttt ggaaatatca ttctttgaag atgcctgctt gcatgcctgt
     5341 cgacactcca aggcaatagc cagttcacag cagcctaagc caacccagcc atcagacttc
     5401 ttaaacacac ctggcaaaga tgaattcata caattccaaa tttcaaccat tgtctctggg
     5461 caataatctg gggctcgctg caacagatgt tttagtcggg attcactttt tgaggagaaa
     5521 atctgttcac atacatcacg gcacatttgg ttatcctttg aatgattaca acacaatgca
     5581 cccgcactgc ccggagccag gccccctgcc acctcggccg gctgggccgc gtcaccagtg
     5641 gaacctggaa cccagagcag cagtacccat agcaggagtg tgtctgtctc catggtggct
     5701 agccagcttg ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg
     5761 gttctctagt tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     5821 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     5881 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     5941 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     6001 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     6061 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     6121 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     6181 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     6241 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     6301 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     6361 gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca
     6421 gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc
     6481 actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat
     6541 cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac
     6601 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6661 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7261 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8401 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8461 ctttgagtga gctgataccg ctcgccgcag ccgaacga
//
