LOCUS       sequence-387077-        7205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD1 with an N-terminal HiBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7205)
  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 7205)
  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-FZD1
FEATURES             Location/Qualifiers
     source          1..7205
                     /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(2796..2813)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2802..3026
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3072..3500
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3159..3178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3369..3390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3509..3529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3514..3843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3694..3829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3756..3775
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3910..4704
                     /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(3964..3983)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4574..4593
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4878..4999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4915..4934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4969..4988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5048..5064)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5048..5064)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5061..5083)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5072..5088
                     /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(5096..5102,5103..5120,5121..5126))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5141..5162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5279..5296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5450..6038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5530..5549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6209..7000,7001..7069))
                     /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     6832..6851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7070..7174)
                     /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 ggatcccagg ctgctggtca gggacctggg caagggcccg ggccggggca
     1081 acaacctcct ccacccccac aacagcagca aagcggacaa caatataatg gggaacgtgg
     1141 gatctccgtt cctgatcacg ggtactgcca acctatctct attccgctct gcactgatat
     1201 cgcgtacaac caaacaatca tgccaaactt gctgggccat actaatcagg aagatgcagg
     1261 cctcgaagtc catcagttct atccgctcgt caaggttcaa tgttctgccg aactcaagtt
     1321 ctttctttgc tctatgtatg caccagtgtg taccgtactg gagcaagctc tgcccccatg
     1381 ccggtcactc tgtgaacgtg ccagacaagg ttgcgaggca ttgatgaata aattcgggtt
     1441 tcagtggcca gataccctga aatgcgagaa gttccccgtt catggggctg gcgaactttg
     1501 cgtcgggcaa aataccagtg acaaaggcac gccaacaccg tccttgctgc cggagttttg
     1561 gacatcaaac cctcaacatg gtggtggtgg tcatcgcggc ggtttccctg gcggggccgg
     1621 cgcgtccgaa cggggcaaat tctcttgtcc gagggcactt aaggtgccat catatctgaa
     1681 ttatcacttt ctcggggaaa aggattgcgg cgctccgtgt gaacctacga aggtgtatgg
     1741 cctcatgtat tttgggccag aggaacttcg ttttagtcgt acctggattg ggatttggtc
     1801 cgtcctttgc tgcgcctcta ccctgtttac ggttctgacg tacttggtcg atatgcgccg
     1861 gttcagttat ccagaaaggc ccatcatatt cttgtccggc tgctatactg cggtagctgt
     1921 ggcctatatt gctggctttc ttctcgaaga tcgcgttgtc tgtaacgata agtttgctga
     1981 agacggagcc cgcaccgtgg ctcaaggaac caagaaagag ggatgcacca ttctctttat
     2041 gatgctctat ttctttagca tggcgtccag catttggtgg gtcattttgt ccctgacatg
     2101 gtttcttgca gcggggatga agtggggaca tgaagccata gaggcgaaca gtcagtactt
     2161 tcatctggca gcttgggccg tcccagcaat taaaactatc acaatactgg ccctcggcca
     2221 agtcgatgga gacgtcctct ctggggtatg tttcgtaggg ctcaataatg tggatgcgct
     2281 tcgagggttc gtgctggcac ctttgtttgt gtatctgttt attggtacta gtttcctgct
     2341 cgcgggattc gtgagcctgt ttcggatcag gacgattatg aaacatgacg gtactaaaac
     2401 tgaaaagctc gaaaagctga tggtcaggat cggagtgttc agtgtcttgt acacggtacc
     2461 tgccaccatt gtcattgcat gctacttcta cgagcaagca ttcagggatc aatgggaacg
     2521 gtcctgggtg gcacaatcct gcaaatccta tgcaatccca tgtccccatc ttcaggccgg
     2581 tggcggagct ccccctcatc cacctatgag cccggacttc actgtgttca tgattaaata
     2641 cctcatgacc cttattgttg gtatcacaag cgggttttgg atttggtctg gcaaaactct
     2701 caacagctgg cgcaaatttt acacacggct tacaaactca aagcaagggg aaaccaccgt
     2761 ctaatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2821 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2881 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2941 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3001 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3061 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3121 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3181 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3241 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3301 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3361 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3421 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3481 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3661 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3781 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3841 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3901 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3961 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4021 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4081 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4141 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4201 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4261 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4321 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4381 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4441 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4501 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4561 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4621 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4681 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4741 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4801 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4861 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4921 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4981 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5041 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5101 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5161 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5221 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5881 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7081 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7141 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7201 acgtc
//
