LOCUS       sequence-208245-        8423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8423)
  AUTHORS   Habedanck R, Stierhof YD, Wilkinson CJ, Nigg EA
  TITLE     The Polo kinase Plk4 functions in centriole duplication.
  JOURNAL   Nat Cell Biol. 2005 Nov;7(11):1140-6.
  PUBMED    16244668
REFERENCE   2  (bases 1 to 8423)
  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: "Nat Cell 
            Biol."; date: "2005-11"; volume: "7(11)"; pages: "1140-6"
COMMENT     Sequence Label: pcDNA3 Plk4(Sak) D154A (Nigg HR26)
FEATURES             Location/Qualifiers
     source          1..8423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        490..693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     644..664
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     738..757
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        738..756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             798..827
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             831..860
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             864..893
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             897..920
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      2672..2681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3889..3906)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3895..4119
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4165..4593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4252..4271)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4462..4483
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4602..4622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4607..4936
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4787..4922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4849..4868
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5003..5797
                     /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(5057..5076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5667..5686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5971..6092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6008..6027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6062..6081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6141..6157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6141..6157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6154..6176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6165..6181
                     /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(6189..6195,6196..6213,6214..6219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6234..6255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6372..6389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6543..7131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6623..6642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7302..8093,8094..8162))
                     /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     7925..7944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8163..8267)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8342..8361)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
       61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      661 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      721 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      781 aacttaagct taccatggag cagaagctga tctccgagga ggacctgaac gagcagaagc
      841 tgatctccga ggaggacctg aacgagcaga agctgatctc cgaggaggac ctgaacctgg
      901 aggtgctgtt ccagggaccc ggggtaccta ggatcctgac catggcgacc tgcatcgggg
      961 agaagatcga ggattttaaa gttggaaatc tgcttggtaa aggatcattt gctggtgtct
     1021 acagagctga gtccattcac actggtttgg aagttgcaat caaaatgata gataagaaag
     1081 ccatgtacaa agcaggaatg gtacagagag tccaaaatga ggtgaaaata cattgccaat
     1141 tgaaacatcc ctctatcttg gagctttata actattttga agatagcaat tatgtgtatc
     1201 tggtattaga aatgtgccat aatggagaaa tgaacaggta tctaaagaat agagtgaaac
     1261 ccttctcaga aaatgaagct cgacacttca tgcaccagat catcacaggg atgttgtatc
     1321 ttcattctca tggtatacta caccgggacc tcacactttc taacctccta ctgactcgta
     1381 atatgaacat caagattgct gcttttgggc tggcaactca actgaaaatg ccacatgaaa
     1441 agcactatac attatgtgga actcctaact acatttcacc agaaattgcc actcgaagtg
     1501 cacatggcct tgaatctgat gtttggtccc tgggctgtat gttttataca ttacttatcg
     1561 ggagaccacc cttcgacact gacacagtca agaacacatt aaataaagta gtattggcag
     1621 attatgaaat gccatctttt ttgtcaatag aggccaagga ccttattcac cagttacttc
     1681 gtagaaatcc agcagatcgt ttaagtctgt cttcagtatt ggaccatcct tttatgtccc
     1741 gaaattcttc aacaaaaagt aaagatttag gaactgtgga agactcaatt gatagtgggc
     1801 atgccacaat ttctactgca attacagctt cttccagtac cagtataagt ggtagtttat
     1861 ttgacaaaag aagacttttg attggtcagc cactcccaaa taaaatgact gtatttccaa
     1921 agaataaaag ttcaactgat ttttcttctt caggagatgg aaacagtttt tatactcagt
     1981 ggggaaatca agaaaccagt aatagtggaa ggggaagagt aattcaagat gcagaagaaa
     2041 ggccacattc tcgatacctt cgtagagctt attcctctga tagatctggc acttctaata
     2101 gtcagtctca agcaaaaaca tatacaatgg aacgatgtca ctcagcagaa atgctttcag
     2161 tgtccaaaag atcaggagga ggtgaaaatg aagagaggta ctcacccaca gacaacaatg
     2221 ccaacatttt taacttcttt aaagaaaaga catccagtag ttctggatct tttgaaagac
     2281 ctgataacaa tcaagcactc tccaatcatc tttgtccagg aaaaactcct tttccatttg
     2341 cagacccgac acctcagact gaaaccgtac aacagtggtt tgggaatctg caaataaatg
     2401 ctcatttaag aaaaactact gaatatgaca gcatcagccc aaaccgggac ttccagggcc
     2461 atccagattt gcagaaggac acatcaaaaa atgcctggac tgatacaaaa gtcaaaaaga
     2521 actctgatgc ttctgataat gcacattctg taaaacagca aaataccatg aaatatatga
     2581 ctgcacttca cagtaaacct gagataatcc aacaagaatg tgtttttggc tcagatcctc
     2641 tttctgaaca gagcaagact aggggtatgg agccaccatg gggttatcag aatcgtacat
     2701 taagaagcat tacatctccg ttggttgctc acaggttaaa accaatcaga cagaaaacca
     2761 aaaaggctgt ggtgagcata cttgattcag aggaggtgtg tgtggagctt gtaaaggagt
     2821 atgcatctca agaatatgtg aaagaagttc ttcagatatc tagtgatgga aatacgatca
     2881 ctatttatta tccaaatggt ggtagaggtt ttcctcttgc tgatagacca ccctcaccta
     2941 ctgacaacat cagtaggtac agctttgaca atttaccaga aaaatactgg cgaaaatatc
     3001 aatatgcttc caggtttgta cagcttgtaa gatctaaatc tcccaaaatc acttatttta
     3061 caagatatgc taaatgcatt ttgatggaga attctcctgg tgctgatttt gaggtttggt
     3121 tttatgatgg ggtaaaaata cacaaaacag aagatttcat tcaggtgatt gaaaagacag
     3181 ggaagtctta cactttaaaa agtgaaagtg aagttaatag cttgaaagag gagataaaaa
     3241 tgtatatgga ccatgctaat gagggtcatc gtatttgttt agcactggaa tccataattt
     3301 cagaagagga aaggaaaact aggagtgctc cctttttccc aataatcata ggaagaaaac
     3361 ctggtagtac tagttcacct aaggccttat cacctcctcc ttctgtggat tcaaattacc
     3421 caacgagaga gagagcatct ttcaacagaa tggtcatgca tagtgctgct tctccaacac
     3481 aggcaccaat ccttaatccc tctatggtta caaatgaagg acttggtctt acaactacag
     3541 cttctggaac agacatctct tctaatagtc taaaagattg tcttcctaaa tcagcacaac
     3601 ttttgaaatc tgtttttgtg aaaaatgttg gttgggctac acagttaact agtggagctg
     3661 tgtgggttca gtttaatgat gggtcccagt tggttgtgca ggcaggagtg tcttctatca
     3721 gttatacctc accaaatggt caaacaacta ggtatggaga aaatgaaaaa ttaccagact
     3781 acatcaaaca gaaattacag tgtctgtctt ccatcctttt gatgttttct aatccgactc
     3841 ctaattttca tctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     3901 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3961 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4021 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4081 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4141 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4201 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4261 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4321 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4381 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4441 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4501 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4561 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4621 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4681 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4741 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4801 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4861 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4921 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4981 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5041 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5101 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5161 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5221 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5281 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5341 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5401 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5461 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5521 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5581 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5641 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5701 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5761 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5821 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5881 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5941 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     6001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6061 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     6121 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6181 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6241 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6301 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6361 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6421 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6481 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6541 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6601 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6661 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6721 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6781 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6841 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6901 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6961 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7021 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7081 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7141 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7201 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7261 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7321 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7381 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7441 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7501 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7561 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7621 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7681 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7741 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7801 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7861 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7921 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7981 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8041 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8101 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8161 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8221 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8281 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     8341 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8401 aggtcgctga gtagtgcgcg agc
//
