LOCUS       sequence-387092-        7106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD7 with an N-terminal HiBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7106)
  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 7106)
  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-FZD7
FEATURES             Location/Qualifiers
     source          1..7106
                     /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(2697..2714)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2703..2927
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2973..3401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3060..3079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3270..3291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3410..3430)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3415..3744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3595..3730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3657..3676
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3811..4605
                     /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(3865..3884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4475..4494
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4779..4900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4816..4835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4870..4889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4949..4965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4949..4965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4962..4984)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4973..4989
                     /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(4997..5003,5004..5021,5022..5027))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5042..5063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5180..5197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5351..5939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5431..5450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6110..6901,6902..6970))
                     /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     6733..6752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6971..7075)
                     /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 ggatcccagc cgtaccacgg agagaagggc atctccgtgc cggaccacgg
     1081 cttctgccag cccatctcca tcccgctgtg cacggacatc gcctacaacc agaccatcct
     1141 gcccaacctg ctgggccaca cgaaccaaga ggacgcgggc ctcgaggtgc accagttcta
     1201 cccgctggtg aaggtgcagt gttctcccga actccgcttt ttcttatgct ccatgtatgc
     1261 gcccgtgtgc accgtgctcg atcaggccat cccgccgtgt cgttctctgt gcgagcgcgc
     1321 ccgccagggc tgcgaggcgc tcatgaacaa gttcggcttc cagtggcccg agcggctgcg
     1381 ctgcgagaac ttcccggtgc acggtgcggg cgagatctgc gtgggccaga acacgtcgga
     1441 cggctccggg ggcccaggcg gcggccccac tgcctaccct accgcgccct acctgccgga
     1501 cctgcccttc accgcgctgc ccccgggggc ctcagatggc agggggcgtc ccgccttccc
     1561 cttctcatgc ccccgtcagc tcaaggtgcc cccgtacctg ggctaccgct tcctgggtga
     1621 gcgcgattgt ggcgccccgt gcgaaccggg ccgtgccaac ggcctgatgt actttaagga
     1681 ggaggagagg cgcttcgccc gcctctgggt gggcgtgtgg tccgtgctgt gctgcgcctc
     1741 gacgctcttt accgttctca cctacctggt ggacatgcgg cgcttcagct acccagagcg
     1801 gcccatcatc ttcctgtcgg gctgctactt catggtggcc gtggcgcacg tggccggctt
     1861 cctactagag gaccgcgccg tgtgcgtgga gcgcttctcg gacgatggct accgcacggt
     1921 ggcgcagggc accaagaagg agggctgcac catcctcttc atggtgctct acttcttcgg
     1981 catggccagc tccatctggt gggtcattct gtctctcact tggttcctgg cggccggcat
     2041 gaagtggggc cacgaggcca tcgaggccaa ctcgcagtac ttccacctgg ccgcgtgggc
     2101 cgtgcccgcc gtcaagacca tcactatcct ggccatgggc caggtagacg gggacctgct
     2161 gagcggggtg tgctacgttg gcctctccag tgtggacgcg ctgcggggct tcgtgctggc
     2221 gcctctgttc gtctacctct tcataggcac gtccttcttg ctggccggct tcgtgtccct
     2281 cttccgtatc cgcaccatca tgaaacacga cggcaccaag accgagaagc tggagaagct
     2341 catggtgcgc atcggcgtct tcagcgtgct ctacacagtg cccgccacca tcgtcctggc
     2401 ctgctacttc tacgagcagg ccttccgcga gcactgggag cgcacctggc tcctgcagac
     2461 gtgcaagagc tatgccgtgc cctgcccgcc cggccacttc ccgcccatga gccccgactt
     2521 caccgtcttc atgatcaagt acctgatgac catgatcgtc ggcatcacca ctggcttctg
     2581 gatctggtcg ggcaagaccc tgcagtcgtg gcgccgcttc taccacagac ttagccacag
     2641 cagcaagggg gagactgcgg tatagtctag agggcccgtt taaacccgct gatcagcctc
     2701 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2761 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2821 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2881 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2941 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3001 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3061 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3121 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3181 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3241 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3301 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3361 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3421 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3481 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3541 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3601 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3661 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3721 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3781 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3841 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3961 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4321 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4621 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4681 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4741 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4861 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4921 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4981 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5041 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5101 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5161 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5221 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5281 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5521 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5761 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6061 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6121 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6181 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6241 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6301 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6361 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6421 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6481 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6541 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6601 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6661 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6721 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6781 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6841 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6901 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6961 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7021 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7081 ttccccgaaa agtgccacct gacgtc
//
