LOCUS       sequence-399915-        5967 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 5967)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5967)
  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: H_NES 80kDa
FEATURES             Location/Qualifiers
     source          1..5967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        23..326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        327..530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             584..610
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(626..628,629..1339)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(626..628,629..1339)
                     /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(668..689)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(929..948)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1276..1297
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2723..2740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2793..2811)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2807..2854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2880..2899
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2917..3372)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3054..3075)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3266..3285
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3399..3503
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3528..3548)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3572..3782
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3594..3790
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3641..3776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3825..4619
                     /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             3825..4619
                     /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(3879..3898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4489..4508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4807..4826)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4851..4898
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5227..5815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5716..5735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
       61 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      301 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      361 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      421 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      481 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      541 accgtcagat ccgctagcgc taccggtcgc caccatgctt caacttcctc ctcttgagag
      601 acttactctt ggaggcgggg gcagcgtgag caagggcgag gagctgttca ccggggtggt
      661 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      721 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      781 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      841 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      901 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      961 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1021 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1081 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1141 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1201 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1261 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1321 catggacgag ctgtacaaga agcttgctca acaaaatgct ttttatcaag tcttaaatat
     1381 gcctaactta aatgctgatc aacgcaatgg ttttatccaa agccttaaag atgatccaag
     1441 ccaaagtgct aacgttttag gtgaagctca aaaacttaat gactctcaag ctccaaaagc
     1501 tgatgcgcaa caaaataact tcaacaaaga tcaacaaagc gccttctatg aaatcttgaa
     1561 catgcctaac ttaaacgaag cgcaacgtaa cggcttcatt caaagtctta aagacgaccc
     1621 aagccaaagc actaacgttt taggtgaagc taaaaaatta aacgaatctc aagcaccgaa
     1681 agctgataac aatttcaaca aagaacaaca aaatgctttc tatgaaatct tgaatatgcc
     1741 taacttaaac gaagaacaac gcaatggttt catccaaagc ttaaaagatg acccaagcca
     1801 aagtgctaac ctattgtcag aagctaaaaa gttaaatgaa tctcaagcac cgaaagcgga
     1861 taacaaattc aacaaagaac aacaaaatgc tttctatgaa atcttacatt tacctaactt
     1921 aaacgaagaa caacgcaatg gtttcatcca aagcctaaaa gatgacccaa gccaaagcgc
     1981 taacctttta gcagaagcta aaaagctaaa tgatgctcaa gcaccaaaag ctcaacaaaa
     2041 tgctttttat caagtcttaa atatgcctaa cttaaatgct gatcaacgca atggttttat
     2101 ccaaagcctt aaagatgatc caagccaaag tgctaacgtt ttaggtgaag ctcaaaaact
     2161 taatgactct caagctccaa aagctgatgc gcaacaaaat aacttcaaca aagatcaaca
     2221 aagcgccttc tatgaaatct tgaacatgcc taacttaaac gaagcgcaac gtaacggctt
     2281 cattcaaagt cttaaagacg acccaagcca aagcactaac gttttaggtg aagctaaaaa
     2341 attaaacgaa tctcaagcac cgaaagctga taacaatttc aacaaagaac aacaaaatgc
     2401 tttctatgaa atcttgaata tgcctaactt aaacgaagaa caacgcaatg gtttcatcca
     2461 aagcttaaaa gatgacccaa gccaaagtgc taacctattg tcagaagcta aaaagttaaa
     2521 tgaatctcaa gcaccgaaag cggataacaa attcaacaaa gaacaacaaa atgctttcta
     2581 tgaaatctta catttaccta acttaaacga agaacaacgc aatggtttca tccaaagcct
     2641 aaaagatgac ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatgaggc
     2701 tcaagcacca aaataactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2761 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2821 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatg
     2881 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     2941 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3001 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3061 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3121 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3181 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3241 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3301 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3361 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3481 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3541 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3721 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3781 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3841 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3901 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3961 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4021 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4081 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4141 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4201 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4261 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4321 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4381 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4441 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4501 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4561 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4621 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4801 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4861 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4921 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4981 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5041 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5941 agtaatcaat tacggggtca ttagttc
//
