LOCUS       sequence-430008-        6377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus transfer vector to express FLAG-tagged mouse Fzd4 
            (codon-optimized for insect cell expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  AUTHORS   Mahoney JP, Bruguera ES, Vasishtha M, Killingsworth LB, Kyaw S, Weis
            WI
  TITLE     PI(4,5)P(2)-stimulated positive feedback drives the recruitment of 
            Dishevelled to Frizzled in Wnt-beta-catenin signaling.
  JOURNAL   Sci Signal. 2022 Aug 23;15(748):eabo2820. doi: 
            10.1126/scisignal.abo2820. Epub 2022 Aug 23.
  PUBMED    35998232
REFERENCE   2  (bases 1 to 6377)
  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"; journalName: "Sci 
            Signal."; date: "2022-08-23"; volume: "15(748):eabo2820. doi"; 
            pages: " 10.1126/scisignal.abo2820. Epub 2022 Aug 23"
COMMENT     Sequence Label: FLAG-mFzd4-opt(40-537)-9xHis_pFastBac1
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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     complement(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             complement(4330..4341)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5678..5701
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     polyA_signal    5867..6001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5904..5923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5958..5977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6030..6195
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatcgggcg cggatcccgg tccgaagcgc gcggaattcg agctcgcccc gcccccagcc
     4081 agtgcgctta cctgccagac tgcgcgccat gaagacgatc atcgccctga gctacatctt
     4141 ctgcctggtg ttcgccgact acaaggacga tgatgacgcc atggaggagc gccgctgcga
     4201 cccaatccgc attgccatgt gccagaacct gggatacaat gtgaccaaga tgccaaacct
     4261 ggtgggccac gagctgcaga cagatgccga gctgcagctg acaaccttca cccccctgat
     4321 ccagtacggc tgcagctccc agctgcagtt ctttctgtgc agcgtgtatg tgccgatgtg
     4381 caccgagaag atcaacattc caatcggacc atgcggagga atgtgcctgt ccgtgaagcg
     4441 ccgctgcgag ccagtgctgc gcgagttcgg ctttgcctgg cccgataccc tgaattgctc
     4501 gaagttcccc ccgcagaacg atcacaatca tatgtgcatg gagggaccag gagacgagga
     4561 ggtgccgctg ccacacaaga cgccaatcca gcccggagag gagtgccatt cggtgggctc
     4621 caactcggac cagtacattt gggtgaagcg cagcctgaat tgcgtgctga agtgcggata
     4681 cgatgccggc ctgtatagtc gcagcgccaa ggagttcacc gacatttgga tggccgtgtg
     4741 ggccagtctg tgcttcatca gcacgacatt tacagtgctg accttcctga ttgattcgag
     4801 tcgctttagc tacccggagc gcccaatcat tttcctgtcc atgtgctaca acatctattc
     4861 gattgcctat atcgtgcgcc tgacagtggg acgcgagcgc atctcctgcg actttgagga
     4921 ggccgccgag ccagtgctga ttcaggaggg actgaagaat acgggctgcg ccatcatttt
     4981 cctgctgatg tacttctttg gcatggccag ctccatttgg tgggtgatcc tgaccctgac
     5041 gtggtttctg gccgccggac tgaagtgggg acacgaggcc atcgagatgc actcgagtta
     5101 cttccatatt gccgcctggg ccatcccagc cgtgaagacc attgtgatcc tgattatgcg
     5161 cctggtggat gccgacgagc tgacgggact gtgctacgtg ggcaaccaga atctggacgc
     5221 cctgacaggc tttgtggtgg cccccctgtt cacctatctg gtgattggaa cgctgtttat
     5281 cgccgccgga ctggtggccc tgttcaagat ccgcagcaac ctgcagaagg atggaacaaa
     5341 gaccgacaag ctggagcgcc tgatggtgaa gattggcgtg ttctccgtgc tgtacacagt
     5401 gcccgccacc tgcgtgattg cctgctactt ttatgagatc tcgaactggg ccctgttccg
     5461 ctatagtgcc gatgacagca atatggccgt ggagatgctg aagatcttca tgtccctgct
     5521 ggtgggaatt acctccggca tgtggatctg gtcggccaag acgctgcaca catggcagaa
     5581 gtgcagtaac cgcctggtga atagcggaaa ggtgaagcgc gagaagcgcg gcaatggctg
     5641 ggtgaagccc ggaaagggaa acgagaccgt ggtgtaacat catcatcatc atcatcatca
     5701 ttaatctaga gcctgcagtc tcgaggcatg cggtaccaag cttgtcgaga agtactagag
     5761 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     5821 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5881 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5941 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     6001 ctgatcactg cttgagccta ggagatccga accagataag tgaaatctag ttccaaacta
     6061 ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc catctatttt
     6121 gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta
     6181 ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc
     6241 caactccatg tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt
     6301 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca
     6361 gcctgaatgg cgaatgg
//
