LOCUS       sequence-430217-        8001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Importin 5 (Imp5, IPO5) with an N-terminal 
            eGFP fusion. Splice isoform 1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  AUTHORS   Apta-Smith MJ, Hernandez-Fernaud JR, Bowman AJ
  TITLE     Evidence for the nuclear import of histones H3.1 and H4 as monomers.
  JOURNAL   EMBO J. 2018 Oct 1;37(19):e98714. doi: 10.15252/embj.201798714. Epub
            2018 Sep 3.
  PUBMED    30177573
REFERENCE   2  (bases 1 to 8001)
  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: "EMBO J."; 
            date: "2018-10-1"; volume: "37(19):e98714. doi"; pages: " 
            10.15252/embj.201798714. Epub 2018 Sep 3"
COMMENT     Sequence Label: pCL1271
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3287..3408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3324..3343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3378..3397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3415..3870)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3552..3573)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3764..3783
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3897..4001
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4003..4360
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4003..4194
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4026..4046)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4211..4346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4395..5189
                     /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             4395..5189
                     /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(4449..4468)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5059..5078
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5377..5396)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5421..5468
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5797..6385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6286..6305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6560..6863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6864..7067
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7018..7038
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      7106..7115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7112..7114,7115..7117,7118..7828)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7157..7178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7418..7437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tacaacagct gctgaagagg ctagacaaat ggccgccgtt ctcctaagac gtctcttgtc
       61 ctctgcattt gatgaagtct atccagcact tccctctgat gttcagactg ccatcaagag
      121 tgagctactc atgattattc agatggaaac acaatctagc atgaggaaaa aagtttgtga
      181 tattgcggca gaactggcca ggaatttaat agatgaggat ggcaataacc agtggcccga
      241 aggtttgaag ttcctttttg attcagtcag ctctcaaaat gtgggactgc gggaagctgc
      301 ccttcacatt ttctggaact ttcctggaat ttttgggaac cagcaacaac actatttaga
      361 tgtcatcaaa cgaatgttag ttcagtgtat gcaagatcag gaacacccgt cgatcaggac
      421 gttatctgct agagctacag ctgcatttat acttgcaaat gagcataatg ttgctctgtt
      481 caaacatttt gcagacttgc taccgggatt cctacaggcg gtaaatgact cgtgctacca
      541 gaatgatgat tctgtcctaa aatccctcgt tgagattgca gatactgttc caaagtattt
      601 gcgtcctcac ttggaagcaa ctctacagct aagtctaaag ttgtgtggag acactagcct
      661 caacaatatg caacgccagc ttgcccttga agtgatcgtc accctctctg agactgcagc
      721 tgctatgtta agaaaacata ccaatattgt tgcacagact attcctcaga tgttagcaat
      781 gatggttgat ttggaagaag atgaggactg ggcaaatgca gatgaactag aagatgatga
      841 ttttgacagc aatgcagttg caggcgagag tgctctagat cgaatggctt gtggacttgg
      901 tggaaagctc gttctgccga tgatcaagga acacattatg caaatgcttc aaaatcctga
      961 ctggaaatac cggcatgcag gattgatggc cttatctgcc attggtgaag ggtgccacca
     1021 gcaaatggaa ggaattctaa atgagatcgt aaattttgtt ttactttttc tccaggatcc
     1081 tcatccaaga gtaaggtatg cagcctgtaa tgccgtggga cagatggcta cagattttgc
     1141 acctggtttc caaaagaaat ttcatgagaa ggtgattgca gctctgctgc agaccatgga
     1201 agaccaaggc aatcaacgtg tgcaggccca tgcagctgct gccctcatta actttactga
     1261 agactgtccc aagtcactac ttattccata cttggataat ttggtgaaac atctgcattc
     1321 cattatggta ctgaagcttc aagagctgat tcagaaaggc accaagttag ttttggaaca
     1381 agttgtgaca tccattgcat cagttgccga tactgcagaa gaaaaatttg tcccctacta
     1441 tgatttattt atgccatcac tgaagcacat cgttgagaat gcggttcaaa aagaactgag
     1501 acttctgaga ggaaaaacta ttgaatgcat tagcctcatt ggtctggctg ttgggaagga
     1561 aaaattcatg caggatgcat cagatgtgat gcagcttttg ttaaagaccc agacagactt
     1621 caatgatatg gaagatgatg atcctcagat ctcttacatg atctcagcat gggccagaat
     1681 gtgcaaaatc cttggaaaag aatttcagca ataccttcca gtggttatgg ggcctttaat
     1741 gaagacggct tcaattaagc ccgaagtagc ccttttagat acccaagaca tggagaatat
     1801 gagtgatgat gatggttggg aatttgtgaa ccttggagat cagcaaagct ttggtattaa
     1861 aactgcagga ctagaagaaa aatcaactgc ttgccagatg ttggtttgct atgctaagga
     1921 gttaaaggaa ggctttgtgg agtacaccga acaggttgtc aaactgatgg tccctttact
     1981 gaaattttat ttccacgatg gtgttcgagt ggcagcagcg gaatccatgc ctcttctcct
     2041 ggagtgtgca agagtccgtg gtcctgagta tctcacacag atgtggcatt ttatgtgtga
     2101 tgctctaatt aaggccattg gtacagaacc agattcagac gtcctctcag aaataatgca
     2161 ttcttttgca aagtgcattg aagtaatggg agatggatgc cttaataatg aacactttga
     2221 agaactggga ggtatattga aagcaaagct tgaagaacat tttaaaaatc aagaattacg
     2281 acaagttaaa agacaagatg aagactatga tgaacaggtc gaagagtcac tacaagatga
     2341 ggatgataat gatgtttata ttctgaccaa agtgtcagat attttacact caatattcag
     2401 tagctacaaa gaaaaggtgt taccatggtt tgaacagctg cttccattaa ttgtcaacct
     2461 catttgtcca catagaccat ggccagacag acaatgggga ttatgcatct ttgatgatgt
     2521 catagaacac tgtagtccag cctcatttaa atacgcagaa tatttcttaa gaccaatgct
     2581 ccaatatgta tgtgacaaca gcccagaagt caggcaagca gctgcatatg gcctgggagt
     2641 catggcacag tacggtggag ataattatcg ccctttttgt acagaagcac ttcccctgct
     2701 ggtaagagtt attcagtctg cggattctaa gaccaaagaa aatgtcaatg ctacagagaa
     2761 ctgcatctca gcagtaggga aaatcatgaa gttcaagcct gactgtgtaa acgttgaaga
     2821 ggtccttcca cactggttgt cttggcttcc actacatgaa gataaagaag aagctgttca
     2881 gactttcaat tatctgtgtg acctgattga aagtaatcat ccaattgttc ttggcccaaa
     2941 caataccaat ctgcccaaaa tatttagtat aattgcggaa ggagaaatgc acgaggcaat
     3001 taaacatgaa gatccttgtg ccaaacgtct ggccaatgtc gttcgccaag tacagacttc
     3061 tggaggactg tggactgagt gcatagcaca gctcagtcct gagcagcagg ccgccattca
     3121 ggagctcctg aactctgcgt gaggtaccgc gggcccggga tccaccggat ctagataact
     3181 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3241 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     3421 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3481 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3541 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3601 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3661 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3721 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3781 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3841 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3901 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3961 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4021 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4261 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4381 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4441 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4501 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4561 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4621 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4681 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4741 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4801 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4861 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4921 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4981 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5041 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5101 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5161 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5221 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5281 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5341 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5401 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5461 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5521 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5581 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5641 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5701 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5941 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6121 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6181 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6241 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6301 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6361 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6421 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6481 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     6541 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6601 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     6661 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6721 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6781 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6841 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     6901 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6961 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     7021 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     7081 gtcagatccg ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     7141 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     7201 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     7261 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     7321 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     7381 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     7441 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     7501 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     7561 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7621 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     7681 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7741 agaccccaac gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg ccgccgggat
     7801 cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagcaa tggcggcggc
     7861 cgcggcggag cagcaacagt tctacctgct cctgggaaac ctgctcagcc ccgacaatgt
     7921 ggtccggaaa caggcagagg aaacctatga gaatatccca ggccagtcaa agatcacatt
     7981 cctcttacaa gccatcagaa a
//
