LOCUS       sequence-62958-e        7338 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 7338)
  AUTHORS   Liu D, Davydenko O, Lampson MA
  TITLE     Polo-like kinase-1 regulates kinetochore-microtubule dynamics and 
            spindle checkpoint silencing.
  JOURNAL   J Cell Biol. 2012 Aug 20;198(4):491-9. doi: 10.1083/jcb.201205090.
  PUBMED    22908307
REFERENCE   2  (bases 1 to 7338)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2012-08-20- 20"; volume: "198"; 
            issue: "4"; pages: "491-9"
COMMENT     Sequence Label: Plk1 FRET sensor c-jun substrate
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      896..905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(902..904,905..907,908..1618)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(902..904,905..907,908..1618)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(902..904,905..907,908..1618)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(2162..2164,2165..2878)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELYK"
     promoter        complement(2894..2912)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2895..2912)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2932..2949)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2938..3162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3208..3636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3295..3314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3505..3526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3645..3665)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3650..3979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3830..3965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3892..3911
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4046..4840
                     /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(4100..4119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4710..4729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5014..5135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5051..5070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5105..5124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5184..5200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5184..5200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5197..5219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5208..5224
                     /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(5232..5238,5239..5256,5257..5262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5277..5298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5415..5432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5586..6171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5666..5685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6342..7133,7134..7202))
                     /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     6965..6984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7203..7307)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttcgccac
      901 catggtgagc aagggcgagg agctgttcgg cgggatcgtg cccatcctgg tcgagctgga
      961 gggcgacgta aacggccaca ggttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1081 cctcgtgacc accctgacct ggggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1141 gcagcacgac ttcttcaagt ccgtcatgcc cgaaggctac gtccaggagc gtaccatctt
     1201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1321 caagctggag tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa
     1381 cggcatcaag gcccacttca aggcccgcca caacatcacg gacggcagcg tgcagctcgc
     1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgatcctgc ccgacaacca
     1501 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1561 cctgctggag ttcgtgaccg ccgccgggat cactcacggc atggacgagc tgtacaaggc
     1621 tggtaccaag ggtaatggta ggtttttaac tttaaaacca ttgcctgaca gcattattca
     1681 agaaagcctg gagattcagc aaggtgtgaa tccatttttc attggtagat ccgaggattg
     1741 caattgtaaa attgaagaca gtaggttgtc tcgagttcat tgcttcattt tcaaaaagag
     1801 gcatgctgta ggcaaaagca tgtatgaatc tccggcacaa ggtttagatg atatttggta
     1861 ttgccacacc ggaactaacg tgagctattt aaataataac cgcatgatac agggtacgaa
     1921 attcctttta caagacggag atgaaatcaa gatcatttgg gataaaaaca ataaatttgt
     1981 cattggcttt aaagtggaaa ttaacgatac tacaggtctg tttaacgagg gattaggtat
     2041 gttacaagaa caaagagtag tacttaagca aacagccgaa gaaaaagatt tggtgaaaaa
     2101 gttaggagga tccgacgacg ccctgaacgc caccttcctg ccctccgagg gctccggagg
     2161 agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2221 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2281 caagctgacc ctgaagctgc tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2341 cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     2401 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2461 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2521 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2581 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     2641 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     2701 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2761 cctgagctac cagtccgccc tgttcaaaga ccccaacgag aagcgcgatc acatggtcct
     2821 gctggagttc ctgaccgccg ccgggatcac tgaaggcatg aacgagctgt acaagtaatc
     2881 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2941 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3001 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3061 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3121 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3181 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3241 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3301 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3361 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3421 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3481 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3541 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3601 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3661 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3721 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3781 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3841 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3901 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3961 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4021 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4081 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4141 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4201 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4261 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4321 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4381 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4441 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4501 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4561 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4621 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4681 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4741 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4801 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4861 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4921 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4981 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5161 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5221 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5281 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5341 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6481 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7321 aaagtgccac ctgacgtc
//
