LOCUS       sequence-348528-        6700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of Patiria miniata Dishevelled.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  AUTHORS   Swartz SZ, Tan TH, Perillo M, Fakhri N, Wessel GM, Wikramanayake AH,
            Cheeseman IM
  TITLE     Polarized Dishevelled dissolution and reassembly drives embryonic 
            axis specification in sea star oocytes.
  JOURNAL   Curr Biol. 2021 Dec 20;31(24):5633-5641.e4. doi: 
            10.1016/j.cub.2021.10.022. Epub 2021 Nov 4.
  PUBMED    34739818
REFERENCE   2  (bases 1 to 6700)
  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: 
            "10.1016/j.cub.2021.10.022"; journalName: "Curr Biol"; date: 
            "2021-12-20- 20"; volume: "31"; issue: "24"; pages: "5633-5641.e4"
COMMENT     Sequence Label: pZS221
FEATURES             Location/Qualifiers
     source          1..6700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(162..750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(242..261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(921..1712,1713..1781))
                     /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     1544..1563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1782..1886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1912..2367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2049..2070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2261..2280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3336..3354
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3336..3353
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      4664..4673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5407..6120
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5437..5465)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6060..6080
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    6174..6308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6198..6217)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6252..6271
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6421..6441)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6421..6439)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6460..6476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6460..6476)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6473..6495)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6484..6500
                     /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(6508..6514,6515..6532,6533..6538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6553..6574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(6691..6700,1..8))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
       61 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      601 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      841 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      901 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      961 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1021 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1081 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1141 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1201 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1261 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1321 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1381 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1441 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1501 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1561 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1621 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1681 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1741 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1801 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1861 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     1921 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1981 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2041 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2101 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2161 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2221 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2281 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2341 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     2401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc
     2461 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc
     2521 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc
     2581 ggaattggca tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata
     2641 ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat
     2701 tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc
     2761 ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc
     2821 ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct
     2881 attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc
     2941 aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc
     3001 cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc
     3061 agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag
     3121 ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca
     3181 agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac
     3241 gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa
     3301 ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta tagaatacaa
     3361 gctacttgtt ctttttgcag gatccactag tggcgcgcca tggaggaaac gaagataata
     3421 taccacatag acgatgagga caccccgtac cttgtgaagt tgccaattcc tgcggaccaa
     3481 gtgacattag gcgattttaa gaatgttctc aatcggccca actacaaatt cttcttcaag
     3541 tctatggacg atgatttcgg cgttgtcgca gcagaaattg tagaagatga ggccaagctg
     3601 ccatgcttca acggccgggt ggtatcctgg ctcgtcccgg ccgacagcag cacaggagcg
     3661 ccatcggaca cggggtcgca atgccaaagc gaagcaccct cagacatgca gccacccatg
     3721 gaacggacgg gcggcatcgg cgactcgcga ccgccttctt tccatgcgaa tgcaccagga
     3781 agcacagaca aactcagcga cacagaaaca gaatcggtca tcagctcaag aagagggaga
     3841 caccaaagtt caaggaggga cagacatcat aatgattttt ctcgtccgaa gagggaccac
     3901 catcggagcg gtcatgggta cgaaagctcc tccacgttaa tgagcagcga catcgagtct
     3961 acgagttact tcgattccac agacgacgac agcagattta gtagtgcaac agagcgaagc
     4021 agctcatcac gattaatgcg cagcaaacac aggaaacggc ggcggagacc gcgcatgcct
     4081 caagtccagc gtgcctcttc ttttagcagt ataacagact ctactatgtc actcaacatt
     4141 atcacagtaa ccttaaattt ggacaaaatt aacttcttgg gtatcagcat tgttggtcaa
     4201 agcaacaagg gcggggacgg tgggatctac gtgggctcga tcatgaaggg gggtgcggta
     4261 gcagcggacg gcaggatcga gccaggcgac atgatcctgc aggtcaacga agtcagcttt
     4321 gagaacatga gcaacgatga tgccgtcagg gtactacgag aagctgtgca ccagccaggg
     4381 cccatcaaac ttgtcgttgc taagtgctgg gatccttcac ctaagggtta cttcacaatc
     4441 cctaggagtg agccagtccg gccgattgat cccggtgcgt gggtggccca caccaacgcc
     4501 atgaaagtgg ccgcgtacca cggtcaagca ggaccggtca gtccctccat gacctcaatg
     4561 acgtccggca gctcttcact aaccagctct atccccgagt ccgaaagatt ggaggatttc
     4621 ggcccactgt caatcaactc ggacttgaac accatcgtgc gtgccatggc ggccccggac
     4681 tctggcctgg acatcagaga ccgcatgtgg ctcaagatta ccattccaaa cgctttccta
     4741 ggttctgatg ttgtggaatg gctgtacaat cgtgtggagg gtttcacaga tcggcgagat
     4801 gccaggaagt atgcctgcaa cctcctcaag gcgggctaca tccgacacac cgtcaacaag
     4861 atcaccttct ccgagcagtg ctactacatc tttggagacc tgtgcggaaa catggcggcc
     4921 ctgtcgctgg gtgacgacac cccagaagtg gaccaggaca ccctggagcc tctcccccag
     4981 cactggatgc cccccggcca gctaccccca aatataccca cggcccctcc cctcccctac
     5041 cagcccatag gctattccca gtttgacacg gccagctaca ccagttttgg aaacgggagc
     5101 gtaggcagtg ccggaagtgg aggaagcagt ggttcaggcc acagccaaga ccacaaggac
     5161 cggaaaggcc gcagcaaagg cagctctggc agcgaatcgg acgccacctc catcatcagc
     5221 cggtcaacgg cgagagggga cctacccatg cgtgccgacg gctccctgcg gggcgacacg
     5281 ccccctatgt ccgtcaactc catgcccatg ggcgtggcca tccccccgat gggtcaaatg
     5341 acagggctcg cacaggccag cgagattgca agaatgggcc ggccagcgat cgccggacat
     5401 cgattgatga gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
     5461 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
     5521 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
     5581 acacttgtca ctactctcac ttatggtgtt caatgctttt caagataccc agatcatatg
     5641 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
     5701 tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc
     5761 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
     5821 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
     5881 aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta
     5941 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     6001 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
     6061 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
     6121 taagaattca aggcctctcg agcctctaga actatagtga gtcgtattac gtagatccag
     6181 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     6241 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     6301 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     6361 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     6421 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6481 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     6541 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6601 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6661 cggtttgcgt attgggcgct cttccgcttc ctcgctcact
//
