LOCUS       sequence-416926-        6902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid expressing eGFP-dnMCAK driven by CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6902)
  AUTHORS   Li J, Hubisz MJ, Earlie EM, Duran MA, Hong C, Varela AA, Lettera E, 
            Deyell M, Tavora B, Havel JJ, Phyu SM, Amin AD, Budre K, Kamiya E, 
            Cavallo JA, Garris C, Powell S, Reis-Filho JS, Wen H, Bettigole S, 
            Khan AJ, Izar B, Parkes EE, Laughney AM, Bakhoum SF
  TITLE     Non-cell-autonomous cancer progression from chromosomal instability.
  JOURNAL   Nature. 2023 Aug;620(7976):1080-1088. doi: 
            10.1038/s41586-023-06464-z. Epub 2023 Aug 23.
  PUBMED    37612508
REFERENCE   2  (bases 1 to 6902)
  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: 
            "10.1038/s41586-023-06464-z"; journalName: "Nature"; date: 
            "2023-08"; volume: "620"; issue: "7976"; pages: "1080-1088"
COMMENT     Sequence Label: pEGFP-dnMCAK
FEATURES             Location/Qualifiers
     source          1..6902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             159..953
                     /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             159..953
                     /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(213..232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     823..842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1141..1160)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1185..1232
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1561..2149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2050..2069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2324..2627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2628..2831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2782..2802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2870..2879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2876..2878,2879..2881,2882..3592)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2876..2878,2879..2881,2882..3592)
                     /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(2921..2942)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3182..3201)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3529..3550
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5953..6074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5990..6009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6044..6063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6081..6536)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6218..6239)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6430..6449
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6669..6902,1..124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6669..6860
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6692..6712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6877..6902,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
       61 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      121 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      181 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      241 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      301 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
      361 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      421 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      481 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      541 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      601 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      661 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
      721 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
      781 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
      841 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
      901 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
      961 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1021 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1081 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     1141 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     1201 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     1261 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     1321 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     1381 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     1441 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1501 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1561 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1621 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1681 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1741 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1801 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1861 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1921 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1981 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2041 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2101 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2161 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2221 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     2281 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2341 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2401 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2461 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2521 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2581 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     2641 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2701 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2761 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2821 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     2881 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2941 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3001 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3061 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3121 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3181 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3241 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3301 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3361 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3421 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3481 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3541 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     3601 cagatctcga gctcaagctt cgaattcgag gcgcgccatg gagtcgcttc ccgctcgatt
     3661 gtttcctggt ctctccatca agattcaacg cagtaatggc ttaattcaca gtgccaacat
     3721 aagcacagtg aacgtggaga aatcctgtgt ttcagtggaa tggatagaag gaggtaacac
     3781 gaaaggcaaa gagattgatt ttgatgatgt ggctgcaata aacccagaac ttctacagct
     3841 tcttccctta cacccaaagg acaatctgcc actgcaggaa aatgtaacag ttccgaagca
     3901 aaaacgcaga gcagtcaact ccaaaattcc agctccaaaa gaaggtcttc gagcacgtgc
     3961 aactcgcatg gcaactgtcc cagaggttcg aattgccact caggagaatg aaatggaggt
     4021 ggagctgcct gtggctacaa actctcgaaa gcagttttcg gttgccactg gcctccctag
     4081 gccctcctgt cctgcaatga cagaattacc gttgtcgatg gtcagcgagg aggcagaaga
     4141 gcaagtccat cccacccgaa gcacatcttc tgcaaaccct gctcggagga aagcatgtat
     4201 tgtgaaggaa atggaaaaaa tgaagaacaa gcgggaagag aagagggccc agaactcgga
     4261 aataagaata aagcgagctc aggaatatga cagcagcttt ccaaactggg aatttgcccg
     4321 gatgattaaa gaattccggg ttactataga atgtcaccca cttactctga ctgatcctac
     4381 ggaagagcac aggatctgcg tgtgtgtgag gaagcgccca ctgaataagc aagaactggc
     4441 caagaaagaa atcgatgtga tttctgttcc cagcaagtgt ttgctctttg tgcatgaacc
     4501 caaattgaaa gtggacttga caaagtatct ggagaatcag gcattctgct tcgattttgc
     4561 ctttgatgaa acagcttcaa atgaagtcgt ctacaggttc acagcaaggc cactggtaca
     4621 gacgattttt gaagggggaa aagcaacctg ctttgcctat gggcagacgg gcagtgggaa
     4681 gactcacaca atgggtggag acctgtctgg taaatctcag aatacatcta aagggatcta
     4741 tgcaatggct tcccgggatg tcttccttct gaagagtcag cctcgctacc ggaacttaaa
     4801 cttggaagtt tatgtgacat tctttgagat ctacaatggg aaggtgtttg atctgctcaa
     4861 caagaaggcc aagctacgtg tactagaaga cagcaagcag caggtgcagg ttgtgggact
     4921 tcaggagtac ctggttaact gtgctgatga tgtcatcaag atgctcaaca tgggcagtgc
     4981 ctgcaggact tctggacaga cttttgccaa ctccaattct tcccgctccc atgcctgctt
     5041 ccagattctt cttcgagcca aagggagatt gcatggcaaa ttctccttgg tggatctggc
     5101 agggaatgaa cgaggggctg acacttctag tgctgaccgc cagactcgca tggagggtgc
     5161 agagattaac aagtctctcc tagcactgaa ggagtgcatc agggcgctgg gacagaacaa
     5221 ggctgccacc ccattcgctg agagcgcact gactcaggtg ctaagggatt cattcattgg
     5281 ggagaactcg aggacttgca tgattgccat gatctcacca ggcataagct cctgtgaata
     5341 tactttaaac acgctgagat atgcagacag agtcaaggag ctgagccccc acagcgggct
     5401 cagtggagag cagccaattc aaatggaaac ggaagagatg gaagccagct ctaatgggac
     5461 ctcgctggca gtcaatttca aggaagaaga ggagctgtct tcccagatgt ccagctttaa
     5521 tgaagccatg agtcagatca gggagctgga agagagggcc atggaggagc tcagggagat
     5581 catacagcaa ggaccaggct ggcttgagct ctctgagatg actgatcagc cagactatga
     5641 cttagagacc tttgtgaaca aagcagaatc tgccctgact cagcagacta agcacttctc
     5701 agccctgcga gaagtcatca aggccctgcg tgtggccatg caactggaag aacaggctag
     5761 caaacagatg aacagcaaga aacggcacca atgattaatt aactcgacgg taccgcgggc
     5821 ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     5881 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5941 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6001 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6061 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     6121 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6181 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6241 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6301 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6361 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6421 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6481 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     6721 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6781 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6841 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6901 tc
//
