LOCUS       sequence-491943-        7886 bp    DNA     circular SYN 05-NOV-2025
DEFINITION  Expression vector for mouse Fzd4, N-terminal SNAPf tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7886)
  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 7886)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 5, 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: pRK5-SNAPf-Fzd4
FEATURES             Location/Qualifiers
     source          1..7886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2223..2768)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(2937..2956)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2969..2987)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2970..2987)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3197..3221)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(3218..3421)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3247..3267)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3422..3801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3830..3846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3830..3846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3843..3865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3854..3870
                     /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(3878..3884,3885..3902,3903..3908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3923..3944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4061..4078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4232..4820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4312..4331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4991..5782,5783..5851))
                     /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     5614..5633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5852..5956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6024..6042
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6080..6102)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6202..6221
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6238..6693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6375..6396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6587..6606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6891..6913
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6905..6922
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6906..6922
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6942..7299)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6956..7091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7108..7299
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7256..7276
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    7368..7502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7392..7411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7446..7465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      7506..7515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cttttttttt tttgagtcat tagtttaaag tgaagaaaac aggggctgcc tagaattagg
       61 gatgttgaaa tttcaggaga gagaaatcca ctggatacct tacgtgtgtt agtacagtac
      121 ttttgtttgc tcaaacaatt ctgttttacg acaaaggcac tgaaggattt tgacaccacc
      181 tcttgggagt gcgctacaga gttctaaaca gcagacagtg taggccagaa gatgtttatg
      241 ttagaccagc caaactataa aactctgcaa cacgtctccc aacccagcat ttccagtgtt
      301 tatagatcac tttgctatct gcaagaccta gttactattt aaaatttaaa agggttttaa
      361 aaattaatct ttctccttgc atgttatcaa ctcacagttg aaatcccacc cagtaggagc
      421 aaacactatc accctgggct tcaggaatct aggcgggacc tccacggact ccaagctgtg
      481 acctgctaaa gtgatcaaca ttttgatttt tcacagcttt gaaaacctct aactggcttt
      541 tccattttgg accattccaa agtctgcagt gaagccattg gtcttctgat gctgagttgg
      601 gtgagtggtg gtgggctgac gtgggtgtgt gtgggctgaa gtgtttctaa gtttgttcaa
      661 actgagattc cctgcatcac ttccttcaca atgaggaacg aggaagccgg ctagtcttat
      721 accacagtct cgttgccttt tcctggcttc acccaaccat tccccctctt ctctctcttt
      781 accttcccag aattcaccaa tcggttagaa cacttttgcc acgtgtgaag agttttggca
      841 gaccaaatcc acatgcctga agtgatgccc acgagcaaag acataaaaat tttcaacatt
      901 tcaactgcca tgtttgagtc atctgcagaa tatcgaaaga gtgcccagtt tgagatttca
      961 tagaaataac aggcaatcac acaggttgca ggaaccgtgt acagtactga gaagaccccg
     1021 atcttgacca ttagcctttc caacttgtct gtctttgtcc cgtctttttg aagattggac
     1081 cgaattttga ataaggccac caaacccgcc gcaatgaaca gcgttccaat caccaaatac
     1141 gtaaagagag gagccaccac aaagccagtg agggcatcta ggttttggtt cccaacatag
     1201 cacaagccag tcagttcatc ggcatccact agtctcataa tcaagatgac aatggttttc
     1261 actgcgggaa tagcccaggc tgcgatgtgg aaataagaac tgtgcatttc aatggcttca
     1321 tgaccccact tgagtccggc tgccaaaaac caagtgagtg tcagaataac ccaccaaatg
     1381 gagctggcca ttccaaaaaa gtacatcagc aagaaaatta ttgcacatcc tgtgttctta
     1441 agtccttctt ggatgagaac gggctctgcc gcctcttcaa aatcacagga tatcctttcc
     1501 cggcctacag tcagccgaac aatataagca atgctataaa tattatagca catactgaga
     1561 aatatgatgg ggcgctcagg gtaagaaaac ctggatgaat caatcaggaa ggtcagcacg
     1621 gtgaaggtgg tggagatgaa gcagaggctg gcccacacag ccatccaaat atccgtgaac
     1681 tccttagctg agcggctgta caagccagca tcgtagccac acttgagaac acagttcagg
     1741 ctcctcttca cccagatgta ctgatcagaa ttgcttccca cggagtggca ctcttccccg
     1801 ggctggatgg gagtcttgtg gggcaaggga acctcttcat cacctggtcc ttccatgcac
     1861 atgtggttgt ggtcgttctg gggcgggaac ttgctgcagt tcagggtgtc gggccaggca
     1921 aacccaaatt ctctcaggac tggttcacag cgtctcttga ctgaaaggca catgccaccg
     1981 cacgggccga tggggatgtt gatcttctct gtgcacattg gcacataaac cgaacaaagg
     2041 aagaactgca gctggctgga gcagccgtac tggatgagcg gcgtgaaagt tgtcagctgc
     2101 agctcggcgt ctgtctgcag ctcgtgtccc actaagttgg gcatcttggt cacgttgtag
     2161 ccgaggttct ggcacatggc gatgcggatg gggtcgcagc gccgctcctc ctcgtccccg
     2221 aaacccagcc caggcttgcc cagtctgtgg ccctcgtggg ccagcagcca ctctttcacg
     2281 gcgagcccgc cctcgtagcc ccccacgtcc aggtcgccct gcaccacccg gtggcagggg
     2341 atcagaatgg gcacgggatt tccgctcagg gcggttttca cggcggcggt ggcggcggga
     2401 ttgccggcca gggcggccag gtggctgtag ctgatgacct ctccgaactt caccactttc
     2461 agcagtttcc acagcacctg gcgggtaaag ctctcctgct ggaacactgg gtggtgcagg
     2521 gctggcacag ggaactcctc gatggcctca ggctggtgaa agtaggcgtt gagccaggcg
     2581 gtggcctgca tcagtggctc tggtccgccc agcacggcgg ctggggcagg cacttccacg
     2641 gcgtcggcgg cagatgttcc tttgcccagg aagatgatac ggtgcaggcc ctgttcgcac
     2701 ccagacagtt ccagcttgcc cagagggcta tccagggtgg tgcgcttcat ttcgcagtct
     2761 ttgtccatcc ccagtgtcgg ccgcaggagc aggaggaact gcagcagcag ccccagcctg
     2821 agcccgacgc ctccaggcgc cccccggctg ctcggccctg tgcccggcca ggccatggtg
     2881 gatccccggg gaattcaatc gatagaaccg aggtgcagtt ggacctggga gtggacacct
     2941 gtggagagaa aggcaaagtg gatgttattc tatagtgtca cctaaatcgt atgtgtatga
     3001 tacataaggt tatgtattaa ttgtagccgc gttctaacga agccaagggg gtgggcctat
     3061 agactctata ggcggtactt acgtcactct tggcacgggg aatccgcgtt ccaatgcacc
     3121 gttcccggcc gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg
     3181 gatggcgtct ccaggcgatc tgacggttca ctaaacgagc tctgcttata tagacctccc
     3241 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     3301 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     3361 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     3421 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     3481 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     3541 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     3601 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     3661 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     3721 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     3781 aactagtcaa taatcaatgt cgggcgagct cgaattaatt cgtaatcatg tcatagctgt
     3841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6001 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     6061 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6121 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     6181 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa
     6241 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     6361 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6421 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     6481 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6541 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6601 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6661 ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg
     6721 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6781 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6841 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6901 acgttgtaaa acgacggcca gtgccaagct gttaacagct ttttgcaaaa gcctaggcct
     6961 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     7021 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     7081 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     7141 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     7201 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     7261 aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct
     7321 ctttcagagg ttatttcagg ccatggtgct gcgccgaatt aattcccgat ccagacatga
     7381 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     7441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     7501 ttgggccatg gcggccaagc ttctgcaggt cgagtttttt tttttttttt tttttttttt
     7561 tttcatggtc caagaatgtg cttttaattg taattgacac acacaaatta cacagttaat
     7621 ctgtgtgaat gcaaagttgc caagaaattt aacatttcag ttcttccaga ctggcgtctg
     7681 cctagatgca atcacacaaa ctaaaatatt ttaagtccca attgcttaac attgcttaat
     7741 gcaataggtc tctagggtcg gtaaggtaag tcaacttagc aaatctcatt tttctctgaa
     7801 ttaagtgatt gtcagaaagt gaatcacctt cccttttcct ctcattaggc atgctgcagt
     7861 gcaaaaaaaa atcctataaa aataac
//
