LOCUS       sequence-360424-        7038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged FZD4 swapped with FZD5 linker domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  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 7038)
  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_5linker
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             403..420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(443..460)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    449..673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      719..1147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(806..825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1016..1037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1156..1176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1161..1490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1341..1476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1403..1422
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1557..2351
                     /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(1611..1630)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2221..2240
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2527..2648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2564..2583)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2618..2637
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2697..2713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2697..2713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2710..2732)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2721..2737
                     /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(2745..2751,2752..2769,2770..2775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2790..2811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2928..2945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3099..3687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3179..3198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3858..4649,4650..4718))
                     /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     4481..4500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4719..4823)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4898..4917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5089..5468
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5469..5672
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5623..5643
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5717..5736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5717..5735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5768..5812
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             5813..5836
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 gggtttggtg gccttattca aaattcggtc caatcttcaa aaagacggga caaagacaga
       61 caagttggaa aggctaatgg tcaagatcgg ggtcttctca gtactgtaca cggttcctgc
      121 aacctgtgtg attgcctgtt atttctatga aatctcaaac tgggcactct ttcgatattc
      181 tgcagatgac tcaaacatgg cagttgaaat gttgaaaatt tttatgtctt tgctcgtggg
      241 catcacttca ggcatgtgga tttggtctgc caaaactctt cacacgtggc aaaagtgttc
      301 taaccgattg gtgaattctg ggaaggtaaa gagagagaag agggggaatg gttgggtgaa
      361 gccaggaaaa ggcaacgaga ctgtggtata gctcgactcg agcatcatca ccatcaccat
      421 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
      481 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
      541 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
      601 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
      661 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
      721 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      781 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      841 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      901 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      961 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1021 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1081 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1141 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcacca ggctccccag
     1201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1321 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1441 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1501 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     1561 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     1621 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     1681 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     1741 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     1801 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     1861 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     1921 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1981 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2041 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     2101 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2161 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2221 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2281 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2341 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     2401 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     2461 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     2521 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2581 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2641 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     2701 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2761 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2941 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3001 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3901 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3961 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4021 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4081 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4141 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4201 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4261 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4321 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4381 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4441 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4501 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4561 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4621 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4681 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4801 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     4861 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     4921 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     4981 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     5041 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
     5101 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5161 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5221 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5281 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5341 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5401 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5461 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     5521 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     5581 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     5641 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
     5701 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct
     5761 tggtaccatg tctgcacttc tgatcctagc tcttgttgga gctgcagttg ctgactacaa
     5821 agacgatgac gacaagcttg cggccgcaga ggagcggcgc tgcgacccca tccgcatcgc
     5881 catgtgccag aacctcggct acaacgtgac caagatgccc aacttagtgg gacacgagct
     5941 gcagacagac gccgagctgc agctgacaac tttcacgccg ctcatccagt acggctgctc
     6001 cagccagctg cagttcttcc tttgttcggt ttatgtgcca atgtgcacag agaagatcaa
     6061 catccccatc ggcccgtgcg gtggcatgtg cctttcagtc aagagacgct gtgaaccagt
     6121 cctgagagaa tttgggtttg cctggcccga caccctgaac tgcagcaagt tcccgcccca
     6181 gaacgaccac aaccacatgt gcatggaagg accaggtagc gaggccacca cggcgccccc
     6241 caggcctttc ccagccaagc ccacccttcc aggcccgcca ggggcgccgg cctcgggggg
     6301 cgaatgcccc gctgggggcc cgttcgtgtg caagtgtcgc gagcccttcg tgcccattct
     6361 gaaggagtca cacccgctct acaacaaggt gcggacgggc caggtgccca actgcgcggt
     6421 accctgtggc tacgatgctg gcttgtacag ccgctcagct aaggagttca cggatatttg
     6481 gatggctgtg tgggccagcc tctgcttcat ctccaccacc ttcaccgtgc tgaccttcct
     6541 gattgattca tccaggtttt cttaccctga gcgccccatc atatttctca gtatgtgcta
     6601 taatatttat agcattgctt atattgttcg gctgactgta ggccgggaaa ggatatcctg
     6661 tgattttgaa gaggcggcag agcccgttct catccaagaa ggacttaaga acacaggatg
     6721 tgcaataatt ttcttgctga tgtacttttt tggaatggcc agctccattt ggtgggttat
     6781 tctgacactc acttggtttt tggcagccgg actcaagtgg ggtcatgaag ccattgaaat
     6841 gcacagttct tatttccaca tcgcagcctg ggctattccc gcagtgaaaa ccattgtcat
     6901 cttgattatg agactagtgg atgccgatga actgactggc ttgtgctatg ttgggaacca
     6961 aaacctagat gccctcactg gctttgtggt ggctcctctc tttacgtatt tggtgattgg
     7021 aacgctgttc attgcggc
//
