LOCUS       sequence-399912-        5958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nucleocytoplasmic transport probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5958)
  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"
COMMENT     Sequence Label: L_NLS 80kDa
FEATURES             Location/Qualifiers
     source          1..5958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     466..486
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      554..563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(602..604,605..1315)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(602..604,605..1315)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(905..924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1252..1273
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2699..2716
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2769..2787)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2783..2830
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2856..2875
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2893..3348)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3030..3051)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3242..3261
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3375..3479
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3504..3524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3548..3758
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3570..3766
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3617..3752
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3801..4595
                     /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             3801..4595
                     /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(3855..3874)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4465..4484
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4783..4802)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4827..4874
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5203..5791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5692..5711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt cggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      541 agcgctaccg gtcgccacca tgggcccaaa aaaggccaga aaagttggag gcgggggcag
      601 cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      721 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      901 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1081 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1201 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagaagct
     1321 tgctcaacaa aatgcttttt atcaagtctt aaatatgcct aacttaaatg ctgatcaacg
     1381 caatggtttt atccaaagcc ttaaagatga tccaagccaa agtgctaacg ttttaggtga
     1441 agctcaaaaa cttaatgact ctcaagctcc aaaagctgat gcgcaacaaa ataacttcaa
     1501 caaagatcaa caaagcgcct tctatgaaat cttgaacatg cctaacttaa acgaagcgca
     1561 acgtaacggc ttcattcaaa gtcttaaaga cgacccaagc caaagcacta acgttttagg
     1621 tgaagctaaa aaattaaacg aatctcaagc accgaaagct gataacaatt tcaacaaaga
     1681 acaacaaaat gctttctatg aaatcttgaa tatgcctaac ttaaacgaag aacaacgcaa
     1741 tggtttcatc caaagcttaa aagatgaccc aagccaaagt gctaacctat tgtcagaagc
     1801 taaaaagtta aatgaatctc aagcaccgaa agcggataac aaattcaaca aagaacaaca
     1861 aaatgctttc tatgaaatct tacatttacc taacttaaac gaagaacaac gcaatggttt
     1921 catccaaagc ctaaaagatg acccaagcca aagcgctaac cttttagcag aagctaaaaa
     1981 gctaaatgat gctcaagcac caaaagctca acaaaatgct ttttatcaag tcttaaatat
     2041 gcctaactta aatgctgatc aacgcaatgg ttttatccaa agccttaaag atgatccaag
     2101 ccaaagtgct aacgttttag gtgaagctca aaaacttaat gactctcaag ctccaaaagc
     2161 tgatgcgcaa caaaataact tcaacaaaga tcaacaaagc gccttctatg aaatcttgaa
     2221 catgcctaac ttaaacgaag cgcaacgtaa cggcttcatt caaagtctta aagacgaccc
     2281 aagccaaagc actaacgttt taggtgaagc taaaaaatta aacgaatctc aagcaccgaa
     2341 agctgataac aatttcaaca aagaacaaca aaatgctttc tatgaaatct tgaatatgcc
     2401 taacttaaac gaagaacaac gcaatggttt catccaaagc ttaaaagatg acccaagcca
     2461 aagtgctaac ctattgtcag aagctaaaaa gttaaatgaa tctcaagcac cgaaagcgga
     2521 taacaaattc aacaaagaac aacaaaatgc tttctatgaa atcttacatt tacctaactt
     2581 aaacgaagaa caacgcaatg gtttcatcca aagcctaaaa gatgacccaa gccaaagcgc
     2641 taacctttta gcagaagcta aaaagctaaa tgatgctcaa gcaccaaaat aactcgagca
     2701 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     2761 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2821 gggttttttg ctgaaaggag gaactatatc cggatgtggt ttgtccaaac tcatcaatgt
     2881 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2941 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3001 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3061 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3121 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3181 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3241 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3301 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3361 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3421 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3481 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3601 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3661 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3721 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3781 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3841 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3901 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3961 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4021 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4081 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4141 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4201 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4261 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4321 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4381 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4441 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4501 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4561 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4621 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4681 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4741 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4801 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4861 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4921 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4981 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5041 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5881 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     5941 ttcatagccc atatatgg
//
