LOCUS       sequence-387089-        7156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD5 with an N-terminal HiBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7156)
  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 7156)
  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-FZD5
FEATURES             Location/Qualifiers
     source          1..7156
                     /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(2748..2765)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2754..2978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3024..3452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3111..3130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3321..3342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3461..3481)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3466..3795
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3646..3781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3708..3727
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3862..4656
                     /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(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4830..4951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4867..4886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4921..4940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5000..5016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5000..5016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5013..5035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5024..5040
                     /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(5048..5054,5055..5072,5073..5078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5093..5114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5231..5248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5402..5989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5482..5501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6160..6951,6952..7020))
                     /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     6783..6802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7021..7125)
                     /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 ggatccgcct ctaaggcccc tgtgtgccaa gagatcaccg tgcctatgtg
     1081 cagaggcatc ggctacaacc tgacacacat gcccaaccag ttcaaccacg acacccagga
     1141 tgaggccgga ctggaagtgc accagttttg gcccctggtg gaaatccagt gtagccccga
     1201 cctgcggttc ttcctgtgct ccatgtacac ccctatctgc ctgcctgact accacaagcc
     1261 tctgcctcca tgcagatccg tgtgcgagag ggccaaagcc ggatgtagcc ctctgatgag
     1321 acagtacggc ttcgcctggc ctgagcggat gagctgtgat agactgcctg tgctgggcag
     1381 agatgccgag gtgctgtgca tggactacaa cagaagcgag gccaccaccg ctccacctag
     1441 accttttcca gccaagccta cactgcctgg acctccaggt gctcctgctt ctggcggaga
     1501 atgtcctgct ggcggacctt tcgtgtgcaa gtgcagagaa cccttcgtgc ccatcctgaa
     1561 agagagccat cctctgtaca acaaagtgcg gaccggccag gtgccaaatt gcgccgttcc
     1621 ttgttaccag cctagcttca gcgccgacga gagaaccttc gccaccttct ggatcggcct
     1681 ttggagcgtg ctgtgcttca tcagcacctc tacaaccgtg gccaccttcc tgatcgacat
     1741 ggaacggttc agataccccg agcggcctat catcttcctg agcgcctgtt acctgtgcgt
     1801 gtccctggga tttctcgtgc ggctggttgt gggacatgcc agcgttgcct gtagcagaga
     1861 gcacaaccac atccactacg agacaacagg ccccgctctg tgcaccatcg tgtttctgct
     1921 ggtgtacttc ttcggcatgg ccagctccat ttggtgggtc atcctgagcc tgacctggtt
     1981 tctggccgct ggaatgaagt ggggcaacga ggccattgcc ggctacgccc agtattttca
     2041 tctggccgcc tggctgatcc ccagcgtgaa gtctattaca gccctggctc tgtccagcgt
     2101 ggacggcgat cctgtggccg gaatctgtta cgtgggcaac cagaacctga acagcctgag
     2161 aggcttcgtg ctgggacctc tggtgctgta tctgctcgtg ggcaccctgt ttttgctggc
     2221 cggctttgtg tccctgttcc ggatcagaag cgtgatcaag caaggcggca ccaagaccga
     2281 caagctggaa aaactgatga tccgcatcgg catcttcacc ctgctgtaca cagtgcccgc
     2341 ctctatcgtg gtggcttgct acctgtacga acagcactac agagagagct gggaagccgc
     2401 tctgacatgt gcctgtcctg gccacgatac aggccagcct agagccaagc cagagtactg
     2461 ggtcctgatg ctgaagtact ttatgtgcct ggtcgtgggc atcaccagcg gcgtttggat
     2521 ttggagcggc aagaccgtgg aaagctggcg gaggttcacc agccggtgct gttgcagacc
     2581 tagaagaggc cacaaatctg gcggcgctat ggccgcaggc gattatcctg aggcttctgc
     2641 cgcactgacc ggcagaacag gacctcctgg acctgccgcc acctatcaca agcaagtgtc
     2701 tctgagccac gtctaatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     2761 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2821 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2881 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2941 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3001 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3061 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3361 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3421 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tcccaattag
     3541 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3601 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3721 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3781 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4801 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4921 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4981 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5941 gttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6001 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6061 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6121 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6181 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6241 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6301 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6361 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6421 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6481 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6541 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6601 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6661 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6721 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6781 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6841 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6901 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6961 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7021 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7081 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7141 agtgccacct gacgtc
//
