LOCUS       sequence-495884-        8335 bp    DNA     circular SYN 19-DEC-2025
DEFINITION  Expression vector for mouse Fzd8, N-terminal 3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8335)
  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 8335)
  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-Fzd8
FEATURES             Location/Qualifiers
     source          1..8335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             932..976
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             977..1042
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3076..3698
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3275..3294)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3722..3742)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3740..4038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4092..4886
                     /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(4146..4165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4756..4775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5538..5672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5575..5594)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5629..5648
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5677..5696)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5678..5696)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5710..5726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5710..5726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5723..5745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5734..5750
                     /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(5758..5764,5765..5782,5783..5788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5803..5824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5941..5958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6112..6700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6192..6211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6871..7662,7663..7731))
                     /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     7494..7513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7732..7836)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7950..7969)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8177..8198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcac catgtctgca cttctgatcc tagctcttgt
      961 tggagctgca gttgctgact acaaagacca tgacggtgat tataaagatc atgacatcga
     1021 ttacaaggat gacgatgaca agcttgcggc ttcggccaag gagctggcgt gccaagagat
     1081 cacggtgccg ttgtgcaaag gcatcggtta caactacact tacatgccca accagttcaa
     1141 ccacgacacg caagatgagg cgggcctaga ggtgcaccag ttttggccgc tggtggagat
     1201 acagtgctcc ccggacctca agttctttct gtgtagcatg tacacgccca tctgcctgga
     1261 ggactacaag aagcctctgc cgccttgtcg ctctgtgtgt gaacgcgcca aggccggctg
     1321 cgcgccgctc atgcgccagt acggctttgc ttggcctgac cgcatgcgct gcgatcggtt
     1381 gccggagcag ggcaacccgg acactctgtg catggactac aaccgcaccg acctcaccac
     1441 ggcagctcca agtccaccta gaagattacc tccaccacct cctccaggtg aacaacctcc
     1501 atctggaagt ggacatagca gacctccagg agctaggcct ccacatcgtg gtggaagcag
     1561 taggggcagc ggagacgcag ctgctgctcc tccttcacgt ggtggaaagg ctaggcctcc
     1621 tggtggtgga gctgctcctt gcgagccggg atgccagtgt agagctccaa tggtgagcgt
     1681 gtccagcgaa cgccacccgc tctacaaccg cgtcaagacc ggccagatcg ccaactgtgc
     1741 gctgccctgc cacaacccct tctttagcca ggatgagcgc gccttcaccg tcttctggat
     1801 cggcctgtgg tcggtgctct gcttcgtctc caccttcgcc actgtctcta ccttcctcat
     1861 cgatatggag cgctttaagt acccggaacg gcccatcata ttcctctccg cctgttacct
     1921 cttcgtgtct gtcgggtacc tggtgcgcct ggtggcagga catgagaaag tggcctgcag
     1981 cggtggagct ccaggtgctg gtggagctgg aggtgcaggt ggagcggcgg cggctggcgc
     2041 aggagcagcg ggagcgggag cgagcagtcc aggtgcgcgc ggagagtacg aggagctggg
     2101 tgcagtcgag cagcatgttc gctatgaaac cactggcccc gcgctgtgca cggtggtctt
     2161 tctccttgtc tacttttttg gcatggccag ctccatctgg tgggtaatcc tgtcgctcac
     2221 gtggttcttg gcagctggca tgaagtgggg taatgaggcc atagcaggct actcgcagta
     2281 cttccacctg gccgcgtggc ttgtgcccag cgtcaagtcc atcgcggtgc tggcgctcag
     2341 ctccgtagac ggcgacccgg tggcgggcat ctgctacgtg ggcaaccaga gccttgacaa
     2401 cctacgcggc tttgtgctgg cgccactggt tatctacctc ttcattggga ctatgtttct
     2461 gttagctggc ttcgtgtcgc tgttccgaat ccgttcagtc atcaagcagc aaggaggccc
     2521 aactaagaca cacaagctag aaaaactcat gatccgcttg ggcctcttca ccgtgctcta
     2581 cacggtgccc gctgccgtcg ttgtcgcctg ccttttctat gagcagcaca accgaccgcg
     2641 ctgggaggcc acgcacaact gcccatgcct tcgggacctg caaccggacc aggctcgcag
     2701 gcccgattac gcggtcttca tgctcaagta cttcatgtgc ctagtagtgg gcatcacatc
     2761 gggcgtgtgg gtctggtccg gcaagactct ggagtcctgg cgcgcgttgt gcactaggtg
     2821 ttgctgggca agcaagggag ctgcagtagg tgcgggtgct ggaggcagcg gccctggggg
     2881 cagtggaccc gggcccggtg gaggtggtgg acacggagga ggagggggat ccctctacag
     2941 cgacgtcagt accggcctga cgtggcggtc tggcacggcc agctctgtat cttaccctaa
     3001 gcaaatgcca ttgtcccagg tctgagaatt catcgataga tctgatatcg gtaccagtcg
     3061 actctagagg atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     3121 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     3181 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     3241 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     3301 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     3361 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     3421 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     3481 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     3541 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     3601 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     3661 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg
     3721 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3781 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3901 ccgcccatcc cgcctccgct aactccgccc agttccgccc attctccgcc ccatggctga
     3961 ctaatttttt ttatttatgc aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4021 gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag acaggatgag
     4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4261 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4921 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4981 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5041 ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag
     5101 gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag
     5161 cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc
     5221 tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc ttgcaggcat
     5281 ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga gcgctggcgc
     5341 ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg taaaaaggcc
     5401 gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc acatttgtag
     5461 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5521 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5581 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5641 aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc gtattaaatt
     5701 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5761 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5821 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5881 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5941 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6001 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6061 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6121 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6181 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6241 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6301 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6361 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6421 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6481 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6541 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6601 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6841 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6901 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6961 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7021 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7081 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7141 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7201 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7261 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7321 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7381 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7441 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7501 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7561 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7621 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7681 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7741 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7801 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7861 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     7921 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     7981 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8041 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     8101 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaagag
     8161 tccactatta aagaacgtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8221 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8281 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aattt
//
