LOCUS       sequence-387098-        7190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD9 with an N-terminal HiBiT; contains P199R; 
            I448V; S524N; R556H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7190)
  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 7190)
  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-FZD9
FEATURES             Location/Qualifiers
     source          1..7190
                     /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(2781..2798)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2787..3011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3057..3485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3144..3163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3354..3375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3494..3514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3499..3828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3679..3814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3741..3760
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3895..4689
                     /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(3949..3968)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4559..4578
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4863..4984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4900..4919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4954..4973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5033..5049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5033..5049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5046..5068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5057..5073
                     /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(5081..5087,5088..5105,5106..5111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5126..5147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5264..5281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5435..6023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5515..5534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6194..6985,6986..7054))
                     /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     6817..6836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7055..7159)
                     /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 ggatccctgg agatcggccg cttcgacccg gagcgcgggc gcggggctgc
     1081 gccgtgccag gcggtggaga tccccatgtg ccgcggcatc ggctacaacc tgacccgcat
     1141 gcccaacctg ctgggccaca cgtcgcaggg cgaggcggct gccgagctag cggagttcgc
     1201 gccgctggtg cagtacggct gccacagcca cctgcgcttc ttcctgtgct cgctctacgc
     1261 gcccatgtgc accgaccagg tctcgacgcc cattcccgcc tgccggccca tgtgcgagca
     1321 ggcgcgcctg cgctgcgcgc ccatcatgga gcagttcaac ttcggctggc cggactcgct
     1381 cgactgcgcc cggctgccca cgcgcaacga cccgcacgcg ctgtgcatgg aggcgcccga
     1441 gaacgccacg gccggccccg cggagcccca caagggcctg ggcatgctgc ccgtggcgcc
     1501 gcggcccgcg cgccctcccg gagacctggg cccgggcgcg ggcggcagtg gcacctgcga
     1561 gaaccgcgag aagttccagt acgtggagaa gagccgctcg tgcgcaccgc gctgcgggcc
     1621 cggcgtcgag gtgttctggt cccggcgcga caaggacttc gcgctggtct ggatggccgt
     1681 gtggtcggcg ctgtgcttct tctccaccgc cttcactgtg ctcaccttct tgctggagcc
     1741 ccaccgcttc cagtaccccg agcgccccat catcttcctc tccatgtgct acaacgtcta
     1801 ctcgctggcc ttcctgatcc gtgcggtggc cggagcgcag agcgtggcct gtgaccagga
     1861 ggcgggcgcg ctctacgtga tccaggaggg cctggagaac acgggctgca cgctggtctt
     1921 cctactgctc tactacttcg gcatggccag ctcgctctgg tgggtggtcc tgacgctcac
     1981 ctggttcctg gctgccggga agaaatgggg ccacgaggcc atcgaggccc acggcagcta
     2041 tttccacatg gctgcctggg gcctgcccgc gctcaagacc atcgtcatcc tgaccctgcg
     2101 caaggtggcg ggtgatgagc tgactgggct ttgctacgtg gccagcacgg atgcagcagc
     2161 gctcacgggc ttcgtgctgg tgcccctctc tggctacctg gtgctgggca gtagtttcct
     2221 cctgaccggc ttcgtggccc tcttccacat ccgcaagatc atgaagacgg gcggcaccaa
     2281 cacagagaag ctggagaagc tcatggtcaa ggtcggggtc ttctccatcc tctacacggt
     2341 gcccgccacc tgcgtcatcg tttgctatgt ctacgaacgc ctcaacatgg acttctggcg
     2401 ccttcgggcc acagagcagc catgcgcagc ggccgcgggg cccggaggcc ggagggactg
     2461 ctcgctgcca gggggctcgg tgcccaccgt ggcggtcttc atgctcaaaa ttttcatgtc
     2521 actggtggtg gggatcacca acggcgtctg ggtgtggagc tccaagactt tccagacctg
     2581 gcagagcctg tgctaccgca agatagcagc tggccgggcc cgggccaagg cctgccacgc
     2641 ccccgggagc tacggacgtg gcacgcactg ccactataag gctcccaccg tggtcttgca
     2701 catgactaag acggacccct ctttggagaa ccccacacac ctcacgtaat ctagagggcc
     2761 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2821 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2881 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2941 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3001 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3061 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3121 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3181 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3241 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3301 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3361 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3421 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3481 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3661 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3721 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3781 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3841 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3901 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3961 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4021 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4081 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4141 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4201 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4261 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4321 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4381 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4441 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4501 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4561 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4621 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4681 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4741 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4801 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4861 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4921 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4981 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5041 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5101 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6241 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6601 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
