LOCUS       sequence-270412-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of human Plk1-WT with N-terminal EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  TITLE     Malumbres and de Carcer unpublished Plk plasmids
REFERENCE   2  (bases 1 to 7759)
  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"
COMMENT     Sequence Label: pBabe-EGFP-Hu_Plk1(WT)
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(1388..1390,1391..1393,1394..2104)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1388..1390,1391..1393,1394..2104)
                     /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(1433..1454)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1694..1713)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2041..2062
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2111..2135
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3988..4012)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4089..4109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4094..4423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4274..4409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4433..5032
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4433..4452)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4929..4949
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5253..5722
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5852..5869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6023..6611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6103..6122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6771..7562,7563..7631))
                     /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     7394..7413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7632..7736)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccgt ttaaacttaa
     1381 gcttacgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1441 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1501 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1561 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1621 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1681 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1741 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1801 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1861 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1921 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1981 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     2041 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2101 caagggtacc acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgag
     2161 tgctgcagtg actgcaggga agctggcacg ggcaccggcc gaccctggga aagccggggt
     2221 ccccggagtt gcagctcccg gagctccggc ggcggctcca ccggcgaaag agatcccgga
     2281 ggtcctagtg gacccacgca gccggcggcg ctatgtgcgg ggccgctttt tgggcaaggg
     2341 cggctttgcc aagtgcttcg agatctcgga cgcggacacc aaggaggtgt tcgcgggcaa
     2401 gattgtgcct aagtctctgc tgctcaagcc gcaccagagg gagaagatgt ccatggaaat
     2461 atccattcac cgcagcctcg cccaccagca cgtcgtagga ttccacggct ttttcgagga
     2521 caacgacttc gtgttcgtgg tgttggagct ctgccgccgg aggtctctcc tggagctgca
     2581 caagaggagg aaagccctga ctgagcctga ggcccgatac tacctacggc aaattgtgct
     2641 tggctgccag tacctgcacc gaaaccgagt tattcatcga gacctcaagc tgggcaacct
     2701 tttcctgaat gaagatctgg aggtgaaaat aggggatttt ggactggcaa ccaaagtcga
     2761 atatgacggg gagaggaaga agaccctgtg tgggactcct aattacatag ctcccgaggt
     2821 gctgagcaag aaagggcaca gtttcgaggt ggatgtgtgg tccattgggt gtatcatgta
     2881 taccttgtta gtgggcaaac caccttttga gacttcttgc ctaaaagaga cctacctccg
     2941 gatcaagaag aatgaataca gtattcccaa gcacatcaac cccgtggccg cctccctcat
     3001 ccagaagatg cttcagacag atcccactgc ccgcccaacc attaacgagc tgcttaatga
     3061 cgagttcttt acttctggct atatccctgc ccgtctcccc atcacctgcc tgaccattcc
     3121 accaaggttt tcgattgctc ccagcagcct ggaccccagc aaccggaagc ccctcacagt
     3181 cctcaataaa ggcttggaga accccctgcc tgagcgtccc cgggaaaaag aagaaccagt
     3241 ggttcgagag acaggtgagg tggtcgactg ccacctcagt gacatgctgc agcagctgca
     3301 cagtgtcaat gcctccaagc cctcggagcg tgggctggtc aggcaagagg aggctgagga
     3361 tcctgcctgc atccccatct tctgggtcag caagtgggtg gactattcgg acaagtacgg
     3421 ccttgggtat cagctctgtg ataacagcgt gggggtgctc ttcaatgact caacacgcct
     3481 catcctctac aatgatggtg acagcctgca gtacatagag cgtgacggca ctgagtccta
     3541 cctcaccgtg agttcccatc ccaactcctt gatgaagaag atcaccctcc ttaaatattt
     3601 ccgcaattac atgagcgagc acttgctgaa ggcaggtgcc aacatcacgc cgcgcgaagg
     3661 tgatgagctc gcccggctgc cctacctacg gacctggttc cgcacccgca gcgccatcat
     3721 cctgcacctc agcaacggca gcgtgcagat caacttcttc caggatcaca ccaagctcat
     3781 cttgtgccca ctgatggcag ccgtgaccta catcgacgag aagcgggact tccgcacata
     3841 ccgcctgagt ctcctggagg agtacggctg ctgcaaggag ctggccagcc ggctccgcta
     3901 cgcccgcact atggtggaca agctgctgag ctcacgctcg gccagcaacc gtctcaaggc
     3961 ctcctaatag caagggtggg cgcgccgacc cagctttctt gtacaaagtg gtctcgagtc
     4021 tagagggccc gtttaaacat atgggcccag atctcgagtt aactgatcag tttaaacgtc
     4081 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4141 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4201 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4261 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4321 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4381 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     4441 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     4501 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     4561 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4621 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4681 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4741 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4801 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4861 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4921 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4981 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     5041 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     5101 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     5161 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     5221 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     5281 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     5341 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     5401 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     5461 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     5521 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     5581 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     5641 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     5701 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     5761 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     5821 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5881 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6721 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6781 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6841 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6901 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6961 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7021 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7081 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7141 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7201 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7261 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7321 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7381 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7441 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7501 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7561 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7621 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7681 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7741 tttccccgaa aagtgccac
//
