LOCUS       sequence-360380-        6976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged FZD4 swapped with FZD3 linker domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6976)
  AUTHORS   Ko SB, Mihara E, Park Y, Roh K, Kang C, Takagi J, Bang I, Choi HJ
  TITLE     Functional role of the Frizzled linker domain in the Wnt signaling 
            pathway.
  JOURNAL   Commun Biol. 2022 May 5;5(1):421. doi: 10.1038/s42003-022-03370-4.
  PUBMED    35513706
REFERENCE   2  (bases 1 to 6976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2022-05-5"; volume: "5(1):421. doi"; pages: " 
            10.1038/s42003-022-03370-4"
COMMENT     Sequence Label: pcDNA FZD4_3linker
FEATURES             Location/Qualifiers
     source          1..6976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             351..368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(391..408)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    397..621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      667..1095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(754..773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     964..985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1104..1124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1109..1438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1289..1424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1351..1370
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1505..2299
                     /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(1559..1578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2169..2188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2475..2596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2512..2531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2566..2585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2645..2661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2645..2661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2658..2680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2669..2685
                     /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(2693..2699,2700..2717,2718..2723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2738..2759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2876..2893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3047..3635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3127..3146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3806..4597,4598..4666))
                     /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     4429..4448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4667..4771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4846..4865)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5037..5416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5417..5620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5571..5591
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5665..5684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5665..5683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5716..5760
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             5761..5784
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ggacaaagac agacaagttg gaaaggctaa tggtcaagat cggggtcttc tcagtactgt
       61 acacggttcc tgcaacctgt gtgattgcct gttatttcta tgaaatctca aactgggcac
      121 tctttcgata ttctgcagat gactcaaaca tggcagttga aatgttgaaa atttttatgt
      181 ctttgctcgt gggcatcact tcaggcatgt ggatttggtc tgccaaaact cttcacacgt
      241 ggcaaaagtg ttctaaccga ttggtgaatt ctgggaaggt aaagagagag aagaggggga
      301 atggttgggt gaagccagga aaaggcaacg agactgtggt atagctcgag catcatcacc
      361 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
      421 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
      481 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
      541 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
      601 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
      661 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      721 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      781 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      841 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      901 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      961 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1021 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1081 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     1141 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1261 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1321 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     1381 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     1441 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     1501 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1561 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1621 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1681 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1741 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1801 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1861 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1921 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1981 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     2041 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     2101 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2161 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     2221 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2281 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     2341 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2401 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     2461 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2521 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2581 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     2641 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2701 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2761 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2821 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2881 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2941 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3001 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3061 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3121 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3181 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3241 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3301 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3361 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3421 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3481 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3541 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3601 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3661 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3721 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3781 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3841 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3901 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3961 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4021 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4081 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4141 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4201 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4261 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4321 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4381 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4441 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4501 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4561 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4621 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4681 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4741 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4801 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     4861 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     4921 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     4981 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     5041 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5101 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5161 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5221 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5281 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5341 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5401 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     5461 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5521 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5581 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     5641 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     5701 gttaagcttg gtaccatgtc tgcacttctg atcctagctc ttgttggagc tgcagttgct
     5761 gactacaaag acgatgacga caagcttgcg gccgcagagg agcggcgctg cgaccccatc
     5821 cgcatcgcca tgtgccagaa cctcggctac aacgtgacca agatgcccaa cttagtggga
     5881 cacgagctgc agacagacgc cgagctgcag ctgacaactt tcacgccgct catccagtac
     5941 ggctgctcca gccagctgca gttcttcctt tgttcggttt atgtgccaat gtgcacagag
     6001 aagatcaaca tccccatcgg cccgtgcggt ggcatgtgcc tttcagtcaa gagacgctgt
     6061 gaaccagtcc tgagagaatt tgggtttgcc tggcccgaca ccctgaactg cagcaagttc
     6121 ccgccccaga acgaccacaa ccacatgtgc atggaaccat atcctcgact tgtggatctg
     6181 aatttagctg gagaaccaac tgaaggagcc ccagtggcag tgcagagaga ctatggtttt
     6241 tggtgtcccc gagagttaaa aattgatcct gatctgggtt attcttttct gcatgtgcgt
     6301 gattgttcac ctccttgtgg ctacgatgct ggcttgtaca gccgctcagc taaggagttc
     6361 acggatattt ggatggctgt gtgggccagc ctctgcttca tctccaccac cttcaccgtg
     6421 ctgaccttcc tgattgattc atccaggttt tcttaccctg agcgccccat catatttctc
     6481 agtatgtgct ataatattta tagcattgct tatattgttc ggctgactgt aggccgggaa
     6541 aggatatcct gtgattttga agaggcggca gagcccgttc tcatccaaga aggacttaag
     6601 aacacaggat gtgcaataat tttcttgctg atgtactttt ttggaatggc cagctccatt
     6661 tggtgggtta ttctgacact cacttggttt ttggcagccg gactcaagtg gggtcatgaa
     6721 gccattgaaa tgcacagttc ttatttccac atcgcagcct gggctattcc cgcagtgaaa
     6781 accattgtca tcttgattat gagactagtg gatgccgatg aactgactgg cttgtgctat
     6841 gttgggaacc aaaacctaga tgccctcact ggctttgtgg tggctcctct ctttacgtat
     6901 ttggtgattg gaacgctgtt cattgcggcg ggtttggtgg ccttattcaa aattcggtcc
     6961 aatcttcaaa aagacg
//
