LOCUS       sequence-416925-        7393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid expressing eGFP-Kif2c driven by CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7393)
  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 7393)
  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-Kif2c
FEATURES             Location/Qualifiers
     source          1..7393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             169..963
                     /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             169..963
                     /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(223..242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     833..852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1151..1170)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1195..1242
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1571..2159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2060..2079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2334..2637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2638..2841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2792..2812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2880..2889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2886..2888,2889..2891,2892..3602)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2886..2888,2889..2891,2892..3602)
                     /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(2931..2952)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3192..3211)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3539..3560
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6454..6575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6491..6510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6545..6564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6582..7037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6719..6740)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6931..6950
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7064..7168
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(7170..7393,1..134)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7170..7361
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7193..7213)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(7378..7393,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
       61 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      121 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      181 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      241 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      301 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
      361 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      421 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      481 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      541 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      601 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      661 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
      721 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      781 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      841 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      901 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      961 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     1021 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1081 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     1141 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1201 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     1261 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     1321 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     1381 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     1441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1681 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     2281 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2341 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2401 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2461 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2521 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2581 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2641 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2701 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2761 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2821 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
     2881 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2941 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3001 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3061 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3121 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3181 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3241 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3301 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3361 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3421 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3481 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3541 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3601 agtactcaga tctcgagctc aagcttcgaa ttcggcacga gctccgaatg gccatggact
     3661 cgtcgcttca ggcccgcctg tttcccggtc tcgctatcaa gatccaacgc agtaatggtt
     3721 taattcacag tgccaatgta aggactgtga acttggagaa atcctgtgtt tcagtggaat
     3781 gggcagaagg aggtgccaca aagggcaaag agattgattt tgatgatgtg gctgcaataa
     3841 acccagaact cttacagctt cttcccttac atccgaagga caatctgccc ttgcaggaaa
     3901 atgtaacaat ccagaaacaa aaacggagat ccgtcaactc caaaattcct gctccaaaag
     3961 aaagtcttcg aagccgctcc actcgcatgt ccactgtctc agagcttcgc atcacggctc
     4021 aggagaatga catggaggtg gagctgcctg cagctgcaaa ctcccgcaag cagttttcag
     4081 ttcctcctgc ccccactagg ccttcctgcc ctgcagtggc tgaaatacca ttgaggatgg
     4141 tcagcgagga gatggaagag caagtccatt ccatccgagg cagctcttct gcaaaccctg
     4201 tgaactcagt tcggaggaaa tcatgtcttg tgaaggaagt ggaaaaaatg aagaacaagc
     4261 gagaagagaa gaaggcccag aactctgaaa tgagaatgaa gagagctcag gagtatgaca
     4321 gtagttttcc aaactgggaa tttgcccgaa tgattaaaga atttcgggct actttggaat
     4381 gtcatccact tactatgact gatcctatcg aagagcacag aatatgtgtc tgtgttagga
     4441 aacgcccact gaataagcaa gaattggcca agaaagaaat tgatgtgatt tccattccta
     4501 gcaagtgtct cctcttggta catgaaccca agttgaaagt ggacttaaca aagtatctgg
     4561 agaaccaagc attctgcttt gactttgcat ttgatgaaac agcttcgaat gaagttgtct
     4621 acaggttcac agcaaggcca ctggtacaga caatctttga aggtggaaaa gcaacttgtt
     4681 ttgcatatgg ccagacagga agtggcaaga cacatactat gggcggagac ctctctggga
     4741 aagcccagaa tgcatccaaa gggatctatg ccatggcctc ccgggacgtc ttcctcctga
     4801 agaatcaacc ctgctaccgg aagttgggcc tggaagtcta tgtgacattc ttcgagatct
     4861 acaatgggaa gctgtttgac ctgctcaaca agaaggccaa gctgcgcgtg ctggaggacg
     4921 gcaagcaaca ggtgcaagtg gtggggctgc aggagcatct ggttaactct gctgatgatg
     4981 tcatcaagat gatcgacatg ggcagcgcct gcagaacctc tgggcagaca tttgccaact
     5041 ccaattcctc ccgctcccac gcgtgcttcc aaattattct tcgagctaaa gggagaatgc
     5101 atggcaagtt ctctttggta gatctggcag ggaatgagcg aggcgcggac acttccagtg
     5161 ctgaccggca gacccgcatg gagggcgcag aaatcaacaa gagtctctta gccctgaagg
     5221 agtgcatcag ggccctggga cagaacaagg ctcacacccc gttccgtgag agcaagctga
     5281 cacaggtgct gagggactcc ttcattgggg agaactctag gacttgcatg attgccacga
     5341 tctcaccagg cataagctcc tgtgaatata ctttaaacac cctgagatat gcagacaggg
     5401 tcaaggagct gagcccccac agtgggccca gtggagagca gttgattcaa atggaaacag
     5461 aagagatgga agcctgctct aacggggcgc tgattccagg caatttatcc aaggaagagg
     5521 aggaactgtc ttcccagatg tccagcttta acgaagccat gactcagatc agggagctgg
     5581 aggagaaggc tatggaagag ctcaaggaga tcatacagca aggaccagac tggcttgagc
     5641 tctctgagat gaccgagcag ccagactatg acctggagac ctttgtgaac aaagcggaat
     5701 ctgctctggc ccagcaagcc aagcatttct cagccctgcg agatgtcatc aaggccttgc
     5761 gcctggccat gcagctggaa gagcaggcta gcagacaaat aagcagcaag aaacggcccc
     5821 agtgacgact gcaaataaaa atctgtttgg tttgacaccc agcctcttcc ctggccctcc
     5881 ccagagaact ttgggtacct ggtgggtcta ggcagggtct gagctgggac aggttctggt
     5941 aaatgccaag tatgggggca tctgggccca gggcagctgg ggagggggtc agagtgacat
     6001 gggacactcc ttttctgttc ctcagttgtc gccctcacga gaggaaggag ctcttagtta
     6061 cccttttgtg ttgcccttct ttccatcaag gggaatgttc tcagcataga gctttctccg
     6121 cagcatcctg cctgcgtgga ctggctgcta atggagagct ccctggggtt gtcctggctc
     6181 tggggagaga gacggagcct ttagtacagc tatctgctgg ctctaaacct tctacgcctt
     6241 tgggccgagc actgaatgtc ttgtacttta aaaaaatgtt tctgagacct ctttctactt
     6301 tactgtctcc ctagagatcc tagaggatcc accggatcta gataactgat cataatcagc
     6361 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     6421 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     6481 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6541 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     6601 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     6661 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     6721 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6781 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6841 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6901 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     6961 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     7021 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7081 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7141 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     7201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     7261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     7321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     7381 ccgcccctaa ctc
//
