LOCUS       sequence-204312-        7322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of GFP-tagged Importin beta in mammalian cells 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7322)
  AUTHORS   Ciciarello M, Mangiacasale R, Thibier C, Guarguaglini G, Marchetti 
            E, Di Fiore B, Lavia P
  TITLE     Importin beta is transported to spindle poles during mitosis and 
            regulates Ran-dependent spindle assembly factors in mammalian cells.
  JOURNAL   J Cell Sci. 2004 Dec 15. 117(Pt 26):6511-22.
  PUBMED    15572412
REFERENCE   2  (bases 1 to 7322)
  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"; journalName: "J Cell Sci.
            2004 Dec 15. 117(Pt 26):6511-22."
COMMENT     Sequence Label: pEGFP-N1 Full length Importin beta
FEATURES             Location/Qualifiers
     source          1..7322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3202..3211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3208..3210,3211..3213,3214..3927)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3208..3210,3211..3213,3214..3924)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3253..3274)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3514..3533)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3861..3882
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4050..4171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4087..4106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4141..4160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4178..4633)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4315..4336)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4527..4546
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4660..4764
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4766..5123
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4766..4957
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4789..4809)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4974..5109
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5158..5952
                     /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             5158..5952
                     /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(5212..5231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5822..5841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6140..6159)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6184..6231
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6560..7148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7049..7068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct catggagctg atcaccattc tcgagaagac cgtgtctccc
      601 gatcggctgg agctggaagc ggcgcagaag ttcctggagc gtgcggccgt ggagaacctg
      661 cccactttcc ttgtggaact gtccagagtg ctggcaaatc caggaaacag tcaggttgcc
      721 agagttgcag ctggtctaca aatcaagaac tctttgacat ctaaagatcc agatatcaag
      781 gcacaatatc agcagaggtg gcttgctatt gatgctaatg ctcgacgaga agtcaagaac
      841 tatgttttgc agacattggg tacagaaact taccggccta gttctgcctc acagtgtgtg
      901 gctggtattg cttgtgcaga gatcccagta aaccagtggc cagaactcat tcctcagctg
      961 gtggccaatg tcacaaaccc caacagcaca gagcacatga aggagtcgac attggaagcc
     1021 atcggttata tttgccaaga tatagaccca gagcagctac aagataaatc caatgagatt
     1081 ctgactgcca taatccaggg gatgaggaaa gaagagccta gtaataatgt gaagctagct
     1141 gctacgaatg cactcctgaa ctcattggag ttcaccaaag caaactttga taaagagtct
     1201 gaaaggcact ttattatgca ggtggtctgt gaagccacac agtgtccaga tacgagggta
     1261 cgagtggctg ctttacagaa tctggtgaag ataatgtcct tatattatca gtacatggag
     1321 acatatatgg gtcctgctct ttttgcaatc acaatcgaag caatgaaaag tgacattgat
     1381 gaggtggctt tacaagggat agaattctgg tccaatgtct gtgatgagga aatggatttg
     1441 gccattgaag cttcagaggc agcagaacaa ggacggcccc ctgagcacac cagcaagttt
     1501 tatgcgaagg gagcactaca gtatctggtt ccaatcctca cacagacact aactaaacag
     1561 gacgaaaatg atgatgacga tgactggaac ccctgcaaag cagcaggggt gtgcctcatg
     1621 cttctggcca cctgctgtga agatgacatt gtcccacatg tcctcccctt cattaaagaa
     1681 cacatcaaga acccagattg gcggtaccgg gatgcagcag tgatggcttt tggttgtatc
     1741 ttggaaggac cagagcccag tcagctcaaa ccactagtta tacaggctat gcccacccta
     1801 atagaattaa tgaaagaccc cagtgtagtt gttcgagata cagctgcatg gactgtaggc
     1861 agaatttgtg agctgcttcc tgaagctgcc atcaatgatg tctacttggc tcccctgcta
     1921 cagtgtctga ttgagggtct cagtgctgaa cccagagtgg cttcaaatgt gtgctgggct
     1981 ttctccagtc tggctgaagc tgcttatgaa gctgcagacg ttgctgatga tcaggaagaa
     2041 ccagctactt actgcttatc ttcttcattt gaactcatag ttcagaagct cctagagact
     2101 acagacagac ctgatggaca ccagaacaac ctgagaagtt ctgcatatga atctctgatg
     2161 gaaattgtga aaaacagtgc caaggattgt tatcctgctg tccagaaaac gactttggtc
     2221 atcatggaac gactgcaaca ggttcttcag atggagtcac atatccagag cacatccgat
     2281 agaatccagt tcaatgacct tcagtcttta ctctgtgcaa ctcttcagaa tgttcttcgg
     2341 aaagtgcaac atcaagatgc tttgcagatc tctgatgtgg ttatggcctc cctgttaagg
     2401 atgttccaaa gcacagctgg gtctggggga gtacaagagg atgccctgat ggcagttagc
     2461 acactggtgg aagtgttggg tggtgaattc ctcaagtaca tggaggcctt taaacccttc
     2521 ctgggcattg gattaaaaaa ttatgctgaa taccaggttt gtttggcagc tgtgggctta
     2581 gtgggagact tgtgccgtgc cctgcaatcc aacatcatac ctttctgtga cgaggtgatg
     2641 cagctgcttc tggaaaattt ggggaatgag aacgtccaca ggtctgtgaa gccgcagatt
     2701 ctgtcagtgt ttggtgatat tgcccttgct attggaggag agtttaaaaa atacttagag
     2761 gttgtattga atactcttca gcaggcctcc caagcccagg tggacaagtc agactatgac
     2821 atggtggatt atctgaatga gctaagggaa agctgcttgg aagcctatac tggaatcgtc
     2881 cagggattaa agggggatca ggagaacgta cacccggatg tgatgctggt acaacccaga
     2941 gtagaattta ttctgtcttt cattgaccac attgctggag atgaggatca cacagatgga
     3001 gtagtagctt gtgctgctgg actaataggg gacttatgta cagcatttgg gaaggatgta
     3061 ctgaaattag tagaagctag gccaatgatc catgaattgt taactgaagg gcggagatcg
     3121 aagactaaca aagcaaaaac ccttgctaca tgggcaacaa aagaactgag gaaactgaag
     3181 aaccaagcgg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     3241 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3301 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     3361 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     3421 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3481 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3541 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3601 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3661 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3721 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3781 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     3841 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3901 ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
     3961 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4021 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4081 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4141 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     4201 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4261 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4321 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4381 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4441 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4501 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4561 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4621 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4681 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4741 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4801 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4861 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4921 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4981 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5041 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5101 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     5161 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5221 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5281 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     5341 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5401 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5461 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5521 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5581 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5641 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     5701 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5761 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5821 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5881 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5941 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6001 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6061 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6121 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     6181 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     6241 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     6301 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     6361 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     6421 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6481 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6541 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6601 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6661 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6721 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6781 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6841 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6901 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6961 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7021 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7081 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7141 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7201 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     7261 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     7321 cg
//
