LOCUS       sequence-81224-e        6849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged Kif2A in mammalian cells under the CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6849)
  AUTHORS   Uehara R, Tsukada Y, Kamasaki T, Poser I, Yoda K, Gerlich DW, 
            Goshima G
  TITLE     Aurora B and Kif2A control microtubule length for assembly of a 
            functional central spindle during anaphase.
  JOURNAL   J Cell Biol. 2013 Aug 19;202(4):623-36. doi: 10.1083/jcb.201302123.
  PUBMED    23960144
REFERENCE   2  (bases 1 to 6849)
  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: "2013-08-19- 19"; volume: "202"; 
            issue: "4"; pages: "623-36"
COMMENT     Sequence Label: pEGFP-Kif2A
FEATURES             Location/Qualifiers
     source          1..6849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3637..3758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3674..3693)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3728..3747
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3765..4220)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3902..3923)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4114..4133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4247..4351
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4353..4710
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4353..4544
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4376..4396)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4561..4696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4745..5539
                     /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             4745..5539
                     /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(4799..4818)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5409..5428
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5727..5746)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5771..5818
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6147..6735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6636..6655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctatggc aacggccaac
     1381 ttcggcaaga tccagatcgg gatttacgtg gagatcaagc gcagcgatgg ccgaatacat
     1441 caagcaatgg taacatcttt aaatgaagat aatgaaagtg taactgttga atggatagaa
     1501 aatggagata caaaaggcaa agagattgac ctggagagca tcttttcact taaccctgac
     1561 cttgttcctg atgaagaaat tgaacccagt ccagaaacac ctccacctcc agcatcctca
     1621 gccaaagtaa acaaaattgt aaagaatcga cggactgtag cttctattaa gaatgaccct
     1681 ccttcaagag ataatagagt ggttggttca gcacgtgcac ggcccagtca atttcctgaa
     1741 cagtcttcct ctgcacaaca gaatggtagt gtttcagata tatctccagt tcaagctgca
     1801 aaaaaggaat ttggaccccc ttcacgtaga aaatctaatt gtgtgaaaga agtagaaaaa
     1861 ctgcaagaaa aacgagagaa aaggagattg caacagcaag aacttagaga aaaaagagcc
     1921 caggacgttg atgctacaaa cccaaattat gaaattatgt gtatgatcag agactttaga
     1981 ggaagtttgg attatagacc attaacaaca gcagatccta ttgatgaaca taggatatgt
     2041 gtgtgtgtaa gaaaacgacc actcaataaa aaagaaactc aaatgaaaga tcttgatgta
     2101 atcacaattc ctagtaaaga tgttgtgatg gtacatgaac caaaacaaaa agtagattta
     2161 acaaggtacc tagaaaacca aacatttcgt tttgattatg cctttgatga ctcagctcct
     2221 aatgagatgg tgtatcgctt cacagctaga ccactagtgg aaactatatt tgaaagggga
     2281 atggctacat gctttgctta tgggcagact ggaagtggaa aaactcatac tatgggtggt
     2341 gacttttcag gaaagaacca agattgttct aaaggaattt atgcattagc agctcgagat
     2401 gtctttttaa tgctaaagaa gccaaactat aagaagctag aacttcaagt atatgcaacc
     2461 ttctttgaaa tttatagtgg aaaggtgttt gacttgctaa acaggaaaac aaaattaaga
     2521 gttctagaag atggaaaaca gcaggttcaa gtggtgggat tacaggaacg ggaggtcaaa
     2581 tgtgttgaag atgtactgaa actcattgac ataggcaaca gttgcagaac atccggtcaa
     2641 acatctgcaa atgcacattc atctcggagc catgcagtgt ttcagattat tcttagaagg
     2701 aaaggaaaac tacatggcaa attttctctc attgatttgg ctggaaatga aagaggagct
     2761 gatacttcca gtgcggacag gcaaactagg cttgaaggtg ctgaaattaa taaaagcctt
     2821 ttagcactca aggagtgcat cagagcctta ggtagaaata aacctcatac tcctttccgt
     2881 gcaagtaaac tcactcaggt gttaagagat tctttcatag gtgaaaactc tcgtacctgc
     2941 atgattgcca caatctctcc aggaatggca tcctgtgaaa atactcttaa tacattaaga
     3001 tatgcaaata gggtcaaaga attgactgta gatccaactg ctgctggtga tgttcgtcca
     3061 ataatgcacc atccaccaaa ccagattgat gacttagaga cacagtgggg tgtggggagt
     3121 tcccctcaga gagatgatct aaaacttctt tgtgaacaaa atgaagaaga agtctctcca
     3181 cagttgttta ctttccacga agctgtttca caaatggtag aaatggaaga acaagttgta
     3241 gaagatcaca gggcagtgtt ccaggaatct attcggtggt tagaagatga aaaggccctc
     3301 ttagagatga ctgaagaagt agattatgat gtcgattcat atgctacaca acttgaagct
     3361 attcttgagc aaaaaataga cattttaact gaactgcggg ataaagtgaa atctttccgt
     3421 gcagctctac aagaggagga acaagccagc aagcaaatca acccgaagag accccgtgcc
     3481 ctttaagtcg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
     3541 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3601 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3661 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3721 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     3781 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3841 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3901 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3961 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4021 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4081 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4141 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4201 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4261 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4321 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     4381 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4501 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4561 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4621 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4681 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4741 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4801 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4861 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4921 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4981 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5041 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5101 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5161 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5221 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     5281 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5341 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5401 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5461 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5521 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5581 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5641 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5701 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5761 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5821 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5881 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5941 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     6001 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6121 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6181 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6241 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6301 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6361 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6421 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6481 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6541 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6601 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6661 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6721 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6781 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6841 gccatgcat
//
