LOCUS       sequence-387091-        7543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD6 with an N-terminal HiBiT; contains M345L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7543)
  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 7543)
  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-FZD6
FEATURES             Location/Qualifiers
     source          1..7543
                     /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(3135..3152)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3141..3365
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3411..3839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3498..3517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3708..3729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3848..3868)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3853..4182
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4033..4168
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4095..4114
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4249..5043
                     /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(4303..4322)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4913..4932
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5217..5338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5254..5273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5308..5327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5387..5403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5387..5403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5400..5422)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5411..5427
                     /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(5435..5441,5442..5459,5460..5465))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5480..5501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5618..5635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5789..6376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5869..5888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6547..7338,7339..7407))
                     /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     7170..7189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7408..7512)
                     /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 gtctcttcac ctgtgaacca attactgttc ccagatgtat
     1081 gaaaatggcc tacaacatga cgtttttccc taatctgatg ggtcattatg accagagtat
     1141 tgccgcggtg gaaatggagc attttcttcc tctcgcaaat ctggaatgtt caccaaacat
     1201 tgaaactttc ctctgcaaag catttgtacc aacctgcata gaacaaattc atgtggttcc
     1261 accttgtcgt aaactttgtg agaaagtata ttctgattgc aaaaaattaa ttgacacttt
     1321 tggaatccga tggcctgagg agcttgaatg tgacagatta caatactgtg atgagactgt
     1381 tcctgtaact tttgatccac acacagaatt tcttggtcct cagaagaaaa cagaacaagt
     1441 ccaaagagac attggatttt ggtgtccaag gcatcttaag acttctgggg gacaaggata
     1501 taagtttctg ggaattgacc agtgtgcgcc tccatgcccc aacatgtatt ttaaaagtga
     1561 tgagctagag tttgcaaaaa gttttattgg aacagtttca atattttgtc tttgtgcaac
     1621 tctgttcaca ttccttactt ttttaattga tgttagaaga ttcagatacc cagagagacc
     1681 aattatatat tactctgtct gttacagcat tgtatctctt atgtacttca ttggattttt
     1741 gctgggcgat agcacagcct gcaataaggc agatgagaag ctagaacttg gtgacactgt
     1801 tgtcctaggc tctcaaaata aggcttgcac cgttttgttc atgcttttgt attttttcac
     1861 aatggctggc actgtgtggt gggtgattct taccattact tggttcttag ctgcaggaag
     1921 aaaatggagt tgtgaagcca tcgagcaaaa agcagtgtgg tttcatgctg ttgcatgggg
     1981 aacaccaggt ttcctgactg ttatgcttct tgctctgaac aaagttgaag gagacaacat
     2041 tagtggagtt tgctttgttg gcctttatga cctggatgct tctcgctact ttgtactctt
     2101 gccactgtgc ctttgtgtgt ttgttgggct ctctcttctt ttagctggca ttatttcctt
     2161 aaatcatgtt cgacaagtca tacaacatga tggccggaac caagaaaaac taaagaaatt
     2221 tatgattcga attggagtct tcagcggctt gtatcttgtg ccattagtga cacttctcgg
     2281 atgttacgtc tatgagcaag tgaacaggat tacctgggag ataacttggg tctctgatca
     2341 ttgtcgtcag taccatatcc catgtcctta tcaggcaaaa gcaaaagctc gaccagaatt
     2401 ggctttattt atgataaaat acctgatgac attaattgtt ggcatctctg ctgtcttctg
     2461 ggttggaagc aaaaagacat gcacagaatg ggctgggttt tttaaacgaa atcgcaagag
     2521 agatccaatc agtgaaagtc gaagagtact acaggaatca tgtgagtttt tcttaaagca
     2581 caattctaaa gttaaacaca aaaagaagca ctataaacca agttcacaca agctgaaggt
     2641 catttccaaa tccatgggaa ccagcacagg agctacagca aatcatggca cttctgcagt
     2701 agcaattact agccatgatt acctaggaca agaaactttg acagaaatcc aaacctcacc
     2761 agaaacatca atgagagagg tgaaagcgga cggagcaagt acccccaggt taagagaaca
     2821 ggactgtggt gaacctgcct cgccagcagc atccatctcc agactctctg gggaacaggt
     2881 cgacgggaag ggccaggcag gcagtgtatc tgaaagtgcg cggagtgaag gaaggattag
     2941 tccaaagagt gatattactg acactggcct ggcacagagc aacaatttgc aggtccccag
     3001 ttcttcagaa ccaagcagcc tcaaaggttc cacatctctg cttgttcacc cagtttcagg
     3061 agtgagaaaa gagcagggag gtggttgtca ttcagatact taatctagag ggcccgttta
     3121 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3181 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3241 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3301 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3361 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3421 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3481 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3541 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3601 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3661 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3721 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3781 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3841 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatcc caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4021 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4081 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4141 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4201 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4261 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4321 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4381 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4441 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4501 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4561 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4621 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4681 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4741 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4801 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4861 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4921 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4981 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5041 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5101 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5161 ccggcaggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5341 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5401 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5461 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5521 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5581 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6301 gcaaacaaac caccgctggt agcggtggtt ttttgtttgc aagcagcaga ttacgcgcag
     6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
