LOCUS       sequence-387087-        7412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD3 with an N-terminal HiBiT; contains I292V and 
            V495I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7412)
  AUTHORS   Kozielewicz P, Shekhani R, Moser S, Bowin CF, Wesslowski J, Davidson
            G, Schulte G
  TITLE     Quantitative Profiling of WNT-3A Binding to All Human Frizzled 
            Paralogues in HEK293 Cells by NanoBiT/BRET Assessments.
  JOURNAL   ACS Pharmacol Transl Sci. 2021 May 11;4(3):1235-1245. doi: 
            10.1021/acsptsci.1c00084. eCollection 2021 Jun 11.
  PUBMED    34151213
REFERENCE   2  (bases 1 to 7412)
  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"; doi: 
            "10.1021/acsptsci.1c00084"; journalName: "ACS Pharmacol Transl Sci";
            date: "2021-05-11- 11"; volume: "4"; issue: "3"; pages: "1235-1245"
COMMENT     Sequence Label: HiBiT-FZD3
FEATURES             Location/Qualifiers
     source          1..7412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3003..3020)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3009..3233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3279..3707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3366..3385)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3576..3597
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3716..3736)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3721..4050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3901..4036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3963..3982
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4117..4911
                     /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(4171..4190)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4781..4800
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5085..5206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5122..5141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5176..5195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5255..5271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5255..5271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5268..5290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5279..5295
                     /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(5303..5309,5310..5327,5328..5333))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5348..5369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5486..5503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5657..6245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5737..5756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6416..7207,7208..7276))
                     /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     7039..7058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7277..7381)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgcggct ctgcatcccg caggtgctgt tggccttgtt
      961 cctttccatg ctgacagggc cgggagaagg cagccgggtg agcggctggc ggctgttcaa
     1021 gaagattagc ggatcccaca gtttgttttc ttgtgaacct ataaccttga ggatgtgcca
     1081 agatttgcct tacaatacta ccttcatgcc taatcttctg aaccattatg accaacagac
     1141 tgcagcttta gcaatggagc ccttccaccc tatggtgaac ctggattgtt ctcgggattt
     1201 tcggccattt ctttgtgcac tctatgcccc tatttgtatg gaatatggac gtgtcacact
     1261 tccctgccgt aggctgtgtc agcgtgccta tagcgagtgt tcaaaactca tggagatgtt
     1321 tggtgtcccg tggcctgaag atatggagtg cagtaggttt ccagattgtg atgagccata
     1381 tccccgactt gtggatttga atttagctgg agaaccaact gaaggagccc cagttgcagt
     1441 gcagagggac tatggttttt ggtgtcccag agagttaaaa attgatcctg atcttggcta
     1501 ttcctttctg cacgtgcgag attgttcgcc accatgtccc aatatgtact tcaggagaga
     1561 agaactgtca tttgctcgct atttcatagg cctgatttca atcatttgcc tctctgccac
     1621 attgtttact tttttaacct ttctaattga cgtcacaaga ttccgttacc ctgaaagacc
     1681 tatcatattt tatgcagtct gctacatgat ggtgtcatta attttcttca ttgggttttt
     1741 gctggaggac cgagtagcct gcaatgcatc tatccctgca cagtataagg cttctacagt
     1801 gacacaagga tctcacaata aggcctgtac catgctcttt atggtactat attttttcac
     1861 tatggctggc agtgtatggt gggtaattct taccatcaca tggtttttag cagctgtgcc
     1921 aaagtggggc agtgaagcta ttgagaagaa agcattgctg tttcatgcca gtgcctgggg
     1981 catccccgga actctaacta tcatcctttt agcgatgaat aaaattgaag gtgacaatat
     2041 tagtggcgtg tgttttgtcg gcctctacga cgttgatgca ttaagatatt tcgttctcgc
     2101 tcccctctgc ctgtatgtgg tagttggggt ttctctcctt ttagccggca ttatatccct
     2161 aaacagagtt cggattgaga tcccattaga aaaggaaaac caagataagt tagtgaagtt
     2221 catgatccgg attggtgttt tcagcattct ctaccttgtg ccactcttgg ttgtaattgg
     2281 atgttacttt tatgagcaag cttaccgcgg catctgggag acaacatgga tccaggaacg
     2341 ctgcagagag tatcacattc catgtccgta ccaggttact cagatgagtc gtccagacct
     2401 gattctcttt ctgatgaagt atctcatggc tctcatagtt gggattccct ctatattttg
     2461 ggttggaagc aaaaagacat gctttgaatg ggccagtttt ttccatgggc gtaggaaaaa
     2521 agagatagtg aatgagagcc ggcaggtgct ccaggaacct gactttgctc agtcactcct
     2581 gagggaccca aatactccaa ttataagaaa atcaagagga acttccactc aagggacatc
     2641 cacacatgct tcttcaactc agctggccat ggtggatgac caaagaagca aagcagggag
     2701 tatccacagc aaagtgagca gctaccatgg cagcctccac aggtcacggg atggcaggta
     2761 cactccctgc agttaccgag gaatggagga gagactacct cacggcagca tgtcacggct
     2821 gacggatcat tccaggcaca gtagttctca tcggctcaac gagcagtccc gacacagcag
     2881 catccgagac ctcagtaaca accccatgac tcacattaca catggcacca gcatgaaccg
     2941 tgttattgag gaggatggaa ccagtgctta atctagaggg cccgtttaaa cccgctgatc
     3001 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3061 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3121 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3181 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3241 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3301 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3361 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3421 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3481 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3541 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3601 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3661 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3721 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3841 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3901 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3961 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4021 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4081 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4141 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4201 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4261 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4321 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4381 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4441 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4501 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4561 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4621 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4681 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4741 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4801 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4861 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4921 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4981 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5041 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5101 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5161 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5221 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5281 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5341 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5401 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5461 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5521 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5581 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5641 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5701 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5761 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5821 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5881 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5941 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6001 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6061 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6121 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6181 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6241 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6301 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6361 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6421 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6481 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6541 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6601 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6661 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6721 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6781 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6841 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6901 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6961 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7021 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7081 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7141 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7201 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7261 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7321 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7381 gcacatttcc ccgaaaagtg ccacctgacg tc
//
