LOCUS       sequence-271801-        6709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid expressing GFP-NLS-Plk1-PBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   McKinley KL, Cheeseman IM
  TITLE     Polo-like kinase 1 licenses CENP-A deposition at centromeres.
  JOURNAL   Cell. 2014 Jul 17;158(2):397-411. doi: 10.1016/j.cell.2014.06.016.
  PUBMED    25036634
REFERENCE   2  (bases 1 to 6709)
  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.cell.2014.06"; journalName: "Cell"; date: "2014-07-17- 
            17"; volume: "158"; issue: "2"; pages: "397-411"
COMMENT     Sequence Label: pKM065
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             60..476
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     406..428
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     444..460
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      511..520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(517..519,520..522,523..1233)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(517..519,520..522,523..1233)
                     /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(562..583)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(823..842)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1170..1191
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1279..1299
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1312..1356
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2412..2432)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2417..2746
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2597..2732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2754..3152
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3321..3790
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3920..3937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4091..4679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4171..4190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4839..5630,5631..5699))
                     /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     5462..5481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5700..5804)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5835..6304
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6369..6568
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg
       61 gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga
      121 tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag
      181 aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca
      241 cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct
      301 acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac
      361 accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc
      421 gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccggcc
      481 ggatcccagt gtggtggtac gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      541 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      601 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      661 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
      721 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      781 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      841 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      901 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      961 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1021 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1081 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1141 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1201 gggatcactc tcggcatgga cgagctgtac aagtccggag gggatcggtg gagctccacc
     1261 ggtggcggcc gctctagaga gaatctttat tttcagggcg ccgccaaatt caaagaaacc
     1321 gctgctgcta aattcgaacg ccagcacatg gacagcggag gtggaggtac tagtctcgag
     1381 cgccggaggt ctctcctgga gctgcacaag aggaggaaat cgattgctcc cagcagcctg
     1441 gaccccagca accggaagcc cctcacagtc ctcaataaag gcttggagaa ccccctgcct
     1501 gagcgtcccc gggaaaaaga agaaccagtg gttcgagaga caggtgaggt ggtcgactgc
     1561 cacctcagtg acatgctgca gcagctgcac agtgtcaatg cctccaagcc ctcggagcgt
     1621 gggctggtca ggcaagagga ggctgaggat cctgcctgca tccccatctt ctgggtcagc
     1681 aagtgggtgg actattcgga caagtacggc cttgggtatc agctctgtga taacagcgtg
     1741 ggggtgctct tcaatgactc aacacgcctc atcctctaca atgatggtga cagcctgcag
     1801 tacatagagc gtgacggcac tgagtcctac ctcaccgtga gttcccatcc caactccttg
     1861 atgaagaaga tcaccctcct taaatatttc cgcaattaca tgagcgagca cttgctgaag
     1921 gcaggtgcca acatcacgcc gcgcgaaggt gatgagctcg cccggctgcc ctacctacgg
     1981 acctggttcc gcacccgcag cgccatcatc ctgcacctca gcaacggcag cgtgcagatc
     2041 aacttcttcc aggatcacac caagctcatc ttgtgcccac tgatggcagc cgtgacctac
     2101 atcgacgaga agcgggactt ccgcacatac cgcctgagtc tcctggagga gtacggctgc
     2161 tgcaaggagc tggccagccg gctccgctac gcccgcacta tggtggacaa gctgctgagc
     2221 tcacgctcgg ccagcaaccg tctcaaggcc tcctaaccgc gggcccggga tccaccggat
     2281 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2341 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattcgc cagcacagtg
     2401 gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2461 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2521 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2581 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2641 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2701 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttcatggcca
     2761 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     2821 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     2881 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     2941 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     3001 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     3061 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     3121 tgctgccctc tggttatgtg tgggagggct aagatcgata aaataaaaga ttttatttag
     3181 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     3241 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     3301 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     3361 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     3421 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     3481 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     3541 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     3601 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     3661 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     3721 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     3781 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca
     3841 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3901 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3961 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4021 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4081 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4141 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4201 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4261 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4321 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4381 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4441 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4501 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4561 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4621 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4681 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4741 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa
     4801 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4861 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4921 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4981 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5041 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5101 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5161 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5221 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5281 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5341 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5401 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5461 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5521 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5581 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5641 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5701 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5761 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5821 gtgccacctg cagcctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5881 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     5941 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     6001 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     6061 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     6121 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     6181 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     6241 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     6301 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac
     6361 cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact
     6421 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt
     6481 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
     6541 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
     6601 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     6661 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgct
//
