LOCUS       sequence-387099-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD10 with an N-terminal HiBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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-FZD10
FEATURES             Location/Qualifiers
     source          1..7163
                     /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(2754..2771)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2760..2984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3030..3458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3117..3136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3327..3348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3467..3487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3472..3801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3652..3787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3714..3733
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3868..4662
                     /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(3922..3941)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4532..4551
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4836..4957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4873..4892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4927..4946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5006..5022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5006..5022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5019..5041)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5030..5046
                     /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(5054..5060,5061..5078,5079..5084))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5099..5120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5237..5254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5408..5996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5488..5507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6167..6958,6959..7027))
                     /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     6790..6809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7028..7132)
                     /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 ggatccatca gctccatgga catggagcgc ccgggcgacg gcaaatgcca
     1081 gcccatcgag atcccgatgt gcaaggacat cggctacaac atgactcgta tgcccaacct
     1141 gatgggccac gagaaccagc gcgaggcagc catccagttg cacgagttcg cgccgctggt
     1201 ggagtacggc tgccacggcc acctccgctt cttcctgtgc tcgctgtacg cgccgatgtg
     1261 caccgagcag gtctctaccc ccatccccgc ctgccgggtc atgtgcgagc aggcccggct
     1321 caagtgctcc ccgattatgg agcagttcaa cttcaagtgg cccgactccc tggactgccg
     1381 gaaactcccc aacaagaacg accccaacta cctgtgcatg gaggcgccca acaacggctc
     1441 ggacgagccc acccggggct cgggcctgtt cccgccgctg ttccggccgc agcggcccca
     1501 cagcgcgcag gagcacccgc tgaaggacgg gggccccggg cgcggcggct gcgacaaccc
     1561 gggcaagttc caccacgtgg agaagagcgc gtcgtgcgcg ccgctctgca cgcccggcgt
     1621 ggacgtgtac tggagccgcg aggacaagcg cttcgcagtg gtctggctgg ccatctgggc
     1681 ggtgctgtgc ttcttctcca gcgccttcac cgtgctcacc ttcctcatcg acccggcccg
     1741 cttccgctac cccgagcgcc ccatcatctt cctctccatg tgctactgcg tctactccgt
     1801 gggctacctc atccgcctct tcgccggcgc cgagagcatc gcctgcgacc gggacagcgg
     1861 ccagctctat gtcatccagg agggactgga gagcaccggc tgcacgctgg tcttcctggt
     1921 cctctactac ttcggcatgg ccagctcgct gtggtgggtg gtcctcacgc tcacctggtt
     1981 cctggccgcc ggcaagaagt ggggccacga ggccatcgaa gccaacagca gctacttcca
     2041 cctggcagcc tgggccatcc cggcggtgaa gaccatcctg atcctggtca tgcgcagggt
     2101 ggcgggggac gagctcaccg gggtctgcta cgtgggcagc atggacgtca acgcgctcac
     2161 cggcttcgtg ctcattcccc tggcctgcta cctggtcatc ggcacgtcct tcatcctctc
     2221 gggcttcgtg gccctgttcc acatccggag ggtgatgaag acgggcggcg agaacacgga
     2281 caagctggag aagctcatgg tgcgtatcgg gctcttctct gtgctgtaca ccgtgccggc
     2341 cacctgtgtg atcgcctgct acttttacga acgcctcaac atggattact ggaagatcct
     2401 ggcggcgcag cacaagtgca aaatgaacaa ccagactaaa acgctggact gcctgatggc
     2461 cgcctccatc cccgccgtgg agatcttcat ggtgaagatc tttatgctgc tggtggtggg
     2521 gatcaccagc gggatgtgga tttggacctc caagactctg cagtcctggc agcaggtgtg
     2581 cagccgtagg ttaaagaaga agagccggag aaaaccggcc agcgtgatca ccagcggtgg
     2641 gatttacaaa aaagcccagc atccccagaa aactcaccac gggaaatatg agatccctgc
     2701 ccagtcgccc acctgcgtgt aatctagagg gcccgtttaa acccgctgat cagcctcgac
     2761 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2821 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2881 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2941 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3001 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3061 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3481 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3661 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3721 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3781 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3841 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3901 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3961 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4021 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4081 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4141 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4201 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4261 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4321 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4381 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4441 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4501 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4561 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4621 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4681 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4741 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4801 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4861 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4921 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4981 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5041 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5101 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5161 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5221 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5281 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5341 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5821 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6121 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6181 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6241 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6301 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6361 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6421 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6481 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6541 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6601 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6661 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6721 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6781 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6841 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6901 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6961 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7021 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7141 cccgaaaagt gccacctgac gtc
//
