LOCUS       sequence-308721-        8528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length zebrafish Pard3 (Gene ID: 403050), tagged with EGFP 
            fluorescent protein and Arabidopsis PIF6 (as in construct Addgene ID
            154913)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8528)
  AUTHORS   Buckley CE, Moore RE, Reade A, Goldberg AR, Weiner OD, Clarke JD
  TITLE     Reversible Optogenetic Control of Subcellular Protein Localization 
            in a Live Vertebrate Embryo.
  JOURNAL   Dev Cell. 2016 Jan 11;36(1):117-26. doi: 
            10.1016/j.devcel.2015.12.011.
  PUBMED    26766447
REFERENCE   2  (bases 1 to 8528)
  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.devcel.2015.12"; journalName: "Dev Cell"; date: 
            "2016-01-11- 11"; volume: "36"; issue: "1"; pages: "117-26"
COMMENT     Sequence Label: pCS2+-Pard3-EGFP-PIF6
FEATURES             Location/Qualifiers
     source          1..8528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3516..3525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3522..3524,3525..3527,3528..4238)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3522..3524,3525..3527,3528..4238)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3567..3588)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3828..3847)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4175..4196
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4580..4714
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4604..4623)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4658..4677
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4827..4847)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4827..4845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4866..4882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4866..4882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4879..4901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4890..4906
                     /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(4914..4920,4921..4938,4939..4944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4959..4980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5097..5114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5268..5856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5348..5367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6027..6818,6819..6887))
                     /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     6650..6669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6888..6992)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7018..7473)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7155..7176)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7367..7386
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7544..8528
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg tgttggaact atgaaagtga
      121 cggtgtgttt tgggagaact cgggtcgtgg ttccgtgcgg agacggaaac attaaagtgc
      181 agtcgctcgt ccagcaggcc gcgatgcggt accggagagc gatcgccaag ggtgaggagt
      241 actgggttca ggtttaccgg ctggaacacg gtgatggagg aatcttggac ctggatgatg
      301 tgctgtgtga tgtggccgat gacaaagaca gattagtggc ggtgtttgat gaacaggagc
      361 ctcatgttgg aggagacggc accagtgcca gctctacagg cacacagtct cctgaactct
      421 actgtggaga accgtccacc tccaccccat tgtctgcctt ccaaccctac ctgccccaca
      481 gcgagataga agtcaccaca tccacactgc gcaccaatat gcctttgcat gttcgccgca
      541 gtagtgatcc ggcattgtta aacctgacgg cgatgtcttt ttctgagccg ggctcgcaac
      601 ccgaagagcc ttccaggaaa aaccccacac gctggtccac cactgctggc ttcctgaaac
      661 cacgcttcgc cactggcacc aacagcctgg agaggaaggg tcgaggtgtg gacacatatc
      721 gtagtcttcc tcgtgatgca gggcagtggt ccaatcagaa agagtttcag cgtgaaaagg
      781 cccgttcttc actcagtgcc aatcatccca tggtggaccg ctggctggag aggcaggagc
      841 aggatgagga ggagaatgga aggatagaac cggtgggacg tgcggacaca tgcttggagc
      901 acatgggagt gagatcattg gatgatattg tgaaattggt ggaagtgtcc aatgacgggg
      961 gtccactcgg catccatgtt gtccccttca gtggtcgaga ccgcaggacg ttgggtctgc
     1021 tggtgaagcg attggagcgt ggtggaaagg ctgatgtcca gggtttgttt caggagaatg
     1081 actgcatcat tcggatcaac aacggagatc tgcgcaacgt tcgctttgag caagctcaga
     1141 acatgtttcg tcaagcaatg cgttctcccg tcatcctgtt ccatgtggtg ccaacctcca
     1201 tgaggtccca gtatgagcag atatcccata atgagcacaa ccccagagcg aacatggacc
     1261 tctcgggacg tttcagccca gactccctca ccaatgatct ggacagcgct gctcaccgat
     1321 tggctcaaca cagaccacag ccacccaata accatctgga tactggctct cccgtccatc
     1381 atcttgttgg ctcatcgggc aaacctccga caggccatac ttcatcaccc caaagaggcc
     1441 taagccccgc ccccacaaca ggctttacta agaaggttgg gcggagactc ggaatccagc
     1501 ttaagaaagg acctgaaggt ctcgggttta gcatcacgtc ccgagacgtt cctcttggag
     1561 ggtctgcacc tatttatgtc aagaacattc tgccaagagg agctgccata caagacggca
     1621 gactgaaagc tggagataga ttgctagagg taaatggggt ggatctgaat gggcggggcc
     1681 aggaggaggt ggtctctctg ctaagggcga cgcccatggg tgggactgtc actctggtgg
     1741 tgctgagaca ggaagagacc tttattccca gagagatgaa cgcagaacca gcaatacaga
     1801 acacaagaga atggaaggta gaagaggagg aattggtctt gactccggat ggtacgcggg
     1861 agtttctgac attagaagtg cctctgaatg attctggctc agcaggatta ggagtcagtg
     1921 tcaagggtaa caggtcaaag gaaagccacg ctgacctggg catcttcgtc aagtctatta
     1981 tcaatggagg agcagcttgc aaggacggaa ggctgcgggt caacgatcag cttattgcag
     2041 tcaatgggga gtcgttatta gggaagacaa accaggatgc catggaaacg ctgcgcaagt
     2101 ccatgtcaac ggaaggcaac aagcgaggga tgattcagct gatcgtggcg agacgcatca
     2161 acaagcgcct cgagggtgag agccgcagca gtcctcgagg gttagagcga acactgagcc
     2221 cgtctcctga tgaccatgag cggcgaattt cacactcact gtatgggatc gagggccttg
     2281 atgacaacct gaggccacaa gccagcacaa ataaccgcaa aatgaatcat tatcagctct
     2341 cgccgacggt gaacatgcct caggacgaca cggtcattat tgaagatgac agaccacacg
     2401 ttctccccat ccagctgtct gaccaatcct cgtccagttc tcatgatgac atgggtttcg
     2461 ccgtggagac acccccgccg ccaccctggg aaccagagct tcccgacagc tcttccagtg
     2521 ctaatgctga ggggcagttt cagagggaag gctttggcag gcagagcatg tctgagaaac
     2581 gcactaagca gtatggagat gcaggccaac tcgacatcat caagacccgc aagtccaaga
     2641 gcatggacct agtggctgat gagattaacc tcacacaatg cacagagaac cacacaggct
     2701 cttccacgcg tgatgtgggc ccgtctctgg gactgaagaa gtccagctct ctggaaagtt
     2761 tacagaccgc agtcgcagag gttacgctca atggtgacat gccgtttcac cggccgcgac
     2821 cacggattat aagaggacga ggctgcaatg agagtttcag agctgccatt gacaaatcct
     2881 acgacaggcc tgcagccaat gaggatgaag aggagtgcat ggacacactg gaggaggaca
     2941 ctgaaggaag ctctcgatcc ggccgggatt cagtttccac cgtggccgat ctcactccac
     3001 tccctgtcac cgagcaacag ctgatcaatg gaaaccagcc tgaaaacgac aagaagaaag
     3061 aaaaaggggg gaaggacaaa aagaagccgg agaaagagaa gggtaaaaca aagaaaggaa
     3121 tgctcaaggg cctcggagag atgttcaggt ttggaaagta ccgtaaggat gagcgattag
     3181 acggtgccaa gtggaaagca gaggaaaccc atgcatctga agaggagact cgcagaatga
     3241 agcaagagca ggagaggatc caggcaaaaa cgcgagagat tcgtcagcgt caggccagag
     3301 agcgagacta tgccgagatc caggacttca gtcgatccac cctgacctca cttcctcccg
     3361 aggagcctcc gtacgcaggc atcggctctc tggagcacgg cgggtaccat cgcattcaca
     3421 ctcctccaga ttcaccatac acacagaagc agaacggacg caatggacac ccctccactt
     3481 cagacaggta cgtcgacggt accgcgggcc cggtcgccac catggtgagc aagggcgagg
     3541 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3601 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3661 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3721 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3781 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3841 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3901 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3961 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4021 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4081 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4141 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4201 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagcct
     4261 ctagagccac catgatgttc cttcctactg attattgttg cagactgagc gaccaggaat
     4321 acatggaact cgtcttcgag aacggacaga tactcgcaaa aggccagagg tcaaatgtta
     4381 gtctccataa tcagcggacg aaaagcatca tggatctgta tgaggccgaa tacaacgaag
     4441 attttatgaa aagtattatc catggagggg gtggcgctat taccaacctg ggagataccc
     4501 aagtggtccc acagtcccac gtagcagccg ctcacgagac caatatgctg gagtccaaca
     4561 aacacgtaga cacgcggtag atccagacat gataagatac attgatgagt ttggacaaac
     4621 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4681 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4741 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     4801 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4921 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     7021 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     7081 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     7141 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     7201 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7261 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7321 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7381 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7441 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     7501 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc gccaattcaa
     7561 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     7621 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     7681 tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt gccaactggg
     7741 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc
     7801 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca
     7861 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac
     7921 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata
     7981 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc
     8041 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg
     8101 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc
     8161 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct
     8221 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc
     8281 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagt
     8341 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg
     8401 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc
     8461 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt
     8521 tagggaac
//
