LOCUS       sequence-160351-        6501 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 6501)
  AUTHORS   Lindon C, Pines J
  TITLE     Ordered proteolysis in anaphase inactivates Plk1 to contribute to 
            proper mitotic exit in human cells.
  JOURNAL   J Cell Biol. 2004 Jan 19;164(2):233-41.
  PUBMED    14734534
REFERENCE   2  (bases 1 to 6501)
  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: "J Cell 
            Biol."; date: "2004-01-19"; volume: "164(2)"; pages: "233-41"
COMMENT     Sequence Label: pEYFP-Plk1(499)
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2925..3046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2962..2981)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3016..3035
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3053..3508)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3190..3211)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3402..3421
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3535..3639
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3641..3998
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3641..3832
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3664..3684)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3849..3984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4033..4827
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4033..4827
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4087..4106)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4697..4716
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5015..5034)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5059..5106
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5435..6023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5924..5943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6198..6501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagtactc agatctcgag ctcaagcttc gaattctaag tgctgcagtg actgcaggga
     1021 agctggcacg ggcaccggcc gaccctggga aagccggggt ccccggagtt gcagctcccg
     1081 gagctccggc ggcggctcca ccggcgaaag agatcccgga ggtcctagtg gacccacgca
     1141 gccggcggcg ctatgtgcgg ggccgctttt tgggcaaggg cggctttgcc aagtgcttcg
     1201 agatctcgga cgcggacacc aaggaggtgt tcgcgggcaa gattgtgcct aagtctctgc
     1261 tgctcaagcc gcaccagagg gagaagatgt ccatggaaat atccattcac cgcagcctcg
     1321 cccaccagca cgtcgtagga ttccacggct ttttcgagga caacgacttc gtgttcgtgg
     1381 tgttggagct ctgccgccgg aggtctctcc tggagctgca caagaggagg aaagccctga
     1441 ctgagcctga ggcccgatac tacctacggc aaattgtgct tggctgccag tacctgcacc
     1501 gaaaccgagt tattcatcga gacctcaagc tgggcaacct tttcctgaat gaagatctgg
     1561 aggtgaaaat aggggatttt ggactggcaa ccaaagtcga atatgacggg gagaggaaga
     1621 agaccctgtg tgggactcct aattacatag ctcccgaggt gctgagcaag aaagggcaca
     1681 gtttcgaggt ggatgtgtgg tccattgggt gtatcatgta taccttgtta gtgggcaaac
     1741 caccttttga gacttcttgc ctaaaagaga cctacctccg gatcaagaag aatgaataca
     1801 gtattcccaa gcacatcaac cccgtggccg cctccctcat ccagaagatg cttcagacag
     1861 atcccactgc ccgcccaacc attaacgagc tgcttaatga cgagttcttt acttctggct
     1921 atatccctgc ccgtctcccc atcacctgcc tgaccattcc accaaggttt tcgattgctc
     1981 ccagcagcct ggaccccagc aaccggaagc ccctcacagt cctcaataaa ggcttggaga
     2041 accccctgcc tgagcgtccc cgggaaaaag aagaaccagt ggttcgagag acaggtgagg
     2101 tggtcgactg ccacctcagt gacatgctgc agcagctgca cagtgtcaat gcctccaagc
     2161 cctcggagcg tgggctggtc aggcaagagg aggctgagga tcctgcctgc atccccatct
     2221 tctgggtcag caagtgggtg gactattcgg acaagtacgg ccttgggtat cagctctgtg
     2281 ataacagcgt gggggtgctc ttcaatgact caacacgcct catcctctac aatgatggtg
     2341 acagcctgca gtacatagag cgtgacggca ctgagtccta cctcaccgtg agttcccatc
     2401 ccaactcctt gatgaagaag atcaccctcc ttaaatattt ccgcaattac atgagcgagc
     2461 acttgctgaa ggcaggtgcc aacatcacgc cgcgcgaagg tgatgagctc gcccggctgc
     2521 cctacctacg gacctggttc cgcacccgca gcgccatcat cctgcacctc agcaacggca
     2581 gcgtgcagat caacttcttc caggatcaca ccaagctcat cttgtgccca ctgatggcag
     2641 ccgtgaccta catcgacgag aagcgggact tccgcacata ccgcctgagt ctcctggagg
     2701 agtacggctg ctgcaaggag ctggccagcc ggctccgcta cgcccgcact atggtggaca
     2761 agctgctgag ctcacgctcg gccagcaacc gtctcaaggc ctcctaagct agataactga
     2821 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2881 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2941 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3001 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3061 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3121 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3181 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3241 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3301 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3361 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3421 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3481 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3541 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3601 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3661 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3721 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3841 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3901 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3961 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4021 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4081 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4141 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4201 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4261 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4321 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4381 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4441 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4501 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4561 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4621 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4681 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4741 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4801 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4861 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4921 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4981 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5041 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5101 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5161 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5221 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5281 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5341 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5401 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5461 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5521 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5581 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5641 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5701 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5761 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5821 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5881 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5941 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6001 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6061 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6121 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     6181 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6241 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6301 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6361 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6421 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6481 ttagtcatcg ctattaccat g
//
