LOCUS       sequence-400570-        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: L_NES 80kDa
FEATURES             Location/Qualifiers
     source          1..5967
                     /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"
     CDS             join(611..613,614..1324)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(611..613,614..1324)
                     /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(653..674)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(914..933)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1261..1282
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2708..2725
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2778..2796)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2792..2839
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2865..2884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2902..3357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3039..3060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3251..3270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3384..3488
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3513..3533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3557..3767
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3579..3775
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3626..3761
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3810..4604
                     /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             3810..4604
                     /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(3864..3883)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4474..4493
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4792..4811)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4836..4883
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5212..5800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5701..5720
                     /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 tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      541 agcgctaccg gtcgccacca tgctttatcc tgagagactt agaagaattc ttactggagg
      601 cgggggcagc gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      661 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      721 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      781 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
      841 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      901 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      961 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1021 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1081 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1141 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1201 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1261 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1321 caagaagctt gctcaacaaa atgcttttta tcaagtctta aatatgccta acttaaatgc
     1381 tgatcaacgc aatggtttta tccaaagcct taaagatgat ccaagccaaa gtgctaacgt
     1441 tttaggtgaa gctcaaaaac ttaatgactc tcaagctcca aaagctgatg cgcaacaaaa
     1501 taacttcaac aaagatcaac aaagcgcctt ctatgaaatc ttgaacatgc ctaacttaaa
     1561 cgaagcgcaa cgtaacggct tcattcaaag tcttaaagac gacccaagcc aaagcactaa
     1621 cgttttaggt gaagctaaaa aattaaacga atctcaagca ccgaaagctg ataacaattt
     1681 caacaaagaa caacaaaatg ctttctatga aatcttgaat atgcctaact taaacgaaga
     1741 acaacgcaat ggtttcatcc aaagcttaaa agatgaccca agccaaagtg ctaacctatt
     1801 gtcagaagct aaaaagttaa atgaatctca agcaccgaaa gcggataaca aattcaacaa
     1861 agaacaacaa aatgctttct atgaaatctt acatttacct aacttaaacg aagaacaacg
     1921 caatggtttc atccaaagcc taaaagatga cccaagccaa agcgctaacc ttttagcaga
     1981 agctaaaaag ctaaatgatg ctcaagcacc aaaagctcaa caaaatgctt tttatcaagt
     2041 cttaaatatg cctaacttaa atgctgatca acgcaatggt tttatccaaa gccttaaaga
     2101 tgatccaagc caaagtgcta acgttttagg tgaagctcaa aaacttaatg actctcaagc
     2161 tccaaaagct gatgcgcaac aaaataactt caacaaagat caacaaagcg ccttctatga
     2221 aatcttgaac atgcctaact taaacgaagc gcaacgtaac ggcttcattc aaagtcttaa
     2281 agacgaccca agccaaagca ctaacgtttt aggtgaagct aaaaaattaa acgaatctca
     2341 agcaccgaaa gctgataaca atttcaacaa agaacaacaa aatgctttct atgaaatctt
     2401 gaatatgcct aacttaaacg aagaacaacg caatggtttc atccaaagct taaaagatga
     2461 cccaagccaa agtgctaacc tattgtcaga agctaaaaag ttaaatgaat ctcaagcacc
     2521 gaaagcggat aacaaattca acaaagaaca acaaaatgct ttctatgaaa tcttacattt
     2581 acctaactta aacgaagaac aacgcaatgg tttcatccaa agcctaaaag atgacccaag
     2641 ccaaagcgct aaccttttag cagaagctaa aaagctaaat gatgctcaag caccaaaata
     2701 actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     2761 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     2821 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatgtggtt tgtccaaact
     2881 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2941 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3001 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3061 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3121 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3181 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3241 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3301 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3481 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3601 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3661 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3721 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3781 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3841 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3901 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3961 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4021 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4081 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4141 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4201 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4261 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4321 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4381 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4441 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4501 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4561 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4621 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4681 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4741 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4801 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4861 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4921 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4981 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5041 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5101 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5161 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5221 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5281 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5341 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5401 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5461 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5521 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5581 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5641 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5701 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5761 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5821 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5881 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     5941 ggtcattagt tcatagccca tatatgg
//
