LOCUS       sequence-269688-        7192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human Plk5 W207F with N-terminal EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7192)
  AUTHORS   de Carcer G, Escobar B, Higuero AM, Garcia L, Anson A, Perez G, 
            Mollejo M, Manning G, Melendez B, Abad-Rodriguez J, Malumbres M
  TITLE     Plk5, a polo box domain-only protein with specific roles in neuron 
            differentiation and glioblastoma suppression.
  JOURNAL   Mol Cell Biol. 2011 Mar;31(6):1225-39. doi: 10.1128/MCB.00607-10. 
            Epub 2011 Jan 18.
  PUBMED    21245385
REFERENCE   2  (bases 1 to 7192)
  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: 
            "10.1128/MCB.00607-10"; journalName: "Mol Cell Biol"; date: 
            "2011-03"; volume: "31"; issue: "6"; pages: "1225-39"
COMMENT     Sequence Label: EGFP-Hu_Plk5(W207F)
FEATURES             Location/Qualifiers
     source          1..7192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1211..1230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1402..1781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1782..1985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1936..1956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2030..2049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2030..2048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2087..2089,2090..2092,2093..2803)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2087..2089,2090..2092,2093..2803)
                     /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(2132..2153)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2393..2412)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2740..2761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2810..2834
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3888..3912)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3950..3967)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3956..4180
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4226..4654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4313..4332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4523..4544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4663..4683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4668..4997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4848..4983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4910..4929
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5064..5858
                     /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(5118..5137)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5728..5747
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6032..6153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6069..6088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6123..6142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6202..6218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6202..6218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6215..6237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6226..6242
                     /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(6250..6256,6257..6274,6275..6280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6295..6316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6433..6450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6604..7192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6684..6703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     1381 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     1441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1741 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1981 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     2041 actataggga gacccaagct ggctagcgtt taaacttaag cttacgatgg tgagcaaggg
     2101 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2161 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2221 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2281 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2341 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2401 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2461 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2521 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2581 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2641 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2701 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2761 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtacca caagtttgta
     2821 caaaaaagca ggctccgcgg ccgccccctt caccgccacc atgtacacgg tgctgactgg
     2881 caccccaccc ttcatggcct cacccctgtc ggagatgtac caaaacatcc gtgagggcca
     2941 ctaccccgaa cccgctcacc tgtctgccaa tgcgcgccgc ctcatcgtgc acctcctagc
     3001 acccaacccg gccgagcggc ccagcctgga ccacctgctg caggacgact tcttcacaca
     3061 gggtttcact ccagaccggc tgccggccca ctcctgccac agtcccccca tcttcgccat
     3121 acccccgcct ctgggcagga tcttccggaa ggtgggccag cggctgctca cccagtgccg
     3181 gccaccctgc cccttcacgc ctaaagaggc ctcgggtcca ggagaaggtg ggccagaccc
     3241 tgactccatg gagtgggacg gcgagagctc cctgtctgcg aaagaggttc cctgcctgga
     3301 aggccccatc cacctggtcg cacaagggac cctgcagagt gacctggccg ggcccgaggg
     3361 gagccggcgg ccagaggtgg aggcggccct cagacacctg cagctgtgcc tggatgtagg
     3421 ccccccggcc acacaggacc ccctgggaga gcagcagccc atcctctggg cccccaaatt
     3481 cgtggattat tccagcaaat acggctttgg ctaccagctc ttggacgggg ggcgcacggg
     3541 acggcaccca catggccctg cgaccccccg gagggagggg accctcccca cacctgtgcc
     3601 acctgctgga cccggcctct gcctcctgcg cttcctggcc tctgagcacg ccctgctgct
     3661 gctgttcagc aatgggatgg tgcaggtgag cttcagtgga gtcccggccc aactggtgct
     3721 gagtggcgag ggtgagggtt tgcagctcac cctctgggag caggggtccc ctggcacctc
     3781 ctactccctg gacgtcccgc agagccacgg ctgcgccccc accaccggac agcaccttca
     3841 ccacgccctc cgcatgctgc agagtatcta gaagggtggg cgcgccgacc cagctttctt
     3901 gtacaaagtg gtctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3961 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4021 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4081 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4141 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4201 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4261 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4321 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4381 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4441 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4501 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4561 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4621 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4681 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4741 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4861 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4921 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4981 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5041 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5101 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5161 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5221 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5281 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5341 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5401 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5461 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5521 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5581 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5641 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5701 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5761 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5821 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5881 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5941 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6001 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6061 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6121 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6181 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aa
//
