LOCUS       sequence-399888-        5964 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 5964)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5964)
  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: M_NES 80kDa
FEATURES             Location/Qualifiers
     source          1..5964
                     /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(608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(608..610,611..1321)
                     /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(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2705..2722
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2775..2793)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2789..2836
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2862..2881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2899..3354)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3036..3057)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3248..3267
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3381..3485
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3510..3530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3554..3764
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3576..3772
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3623..3758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3807..4601
                     /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             3807..4601
                     /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(3861..3880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4471..4490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4789..4808)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4833..4880
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5209..5797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5698..5717
                     /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 tgcttcaaaa aaaacttgaa gaacttgaac ttggaggcgg
      601 gggcagcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gaagcttgct caacaaaatg ctttttatca agtcttaaat atgcctaact taaatgctga
     1381 tcaacgcaat ggttttatcc aaagccttaa agatgatcca agccaaagtg ctaacgtttt
     1441 aggtgaagct caaaaactta atgactctca agctccaaaa gctgatgcgc aacaaaataa
     1501 cttcaacaaa gatcaacaaa gcgccttcta tgaaatcttg aacatgccta acttaaacga
     1561 agcgcaacgt aacggcttca ttcaaagtct taaagacgac ccaagccaaa gcactaacgt
     1621 tttaggtgaa gctaaaaaat taaacgaatc tcaagcaccg aaagctgata acaatttcaa
     1681 caaagaacaa caaaatgctt tctatgaaat cttgaatatg cctaacttaa acgaagaaca
     1741 acgcaatggt ttcatccaaa gcttaaaaga tgacccaagc caaagtgcta acctattgtc
     1801 agaagctaaa aagttaaatg aatctcaagc accgaaagcg gataacaaat tcaacaaaga
     1861 acaacaaaat gctttctatg aaatcttaca tttacctaac ttaaacgaag aacaacgcaa
     1921 tggtttcatc caaagcctaa aagatgaccc aagccaaagc gctaaccttt tagcagaagc
     1981 taaaaagcta aatgatgctc aagcaccaaa agctcaacaa aatgcttttt atcaagtctt
     2041 aaatatgcct aacttaaatg ctgatcaacg caatggtttt atccaaagcc ttaaagatga
     2101 tccaagccaa agtgctaacg ttttaggtga agctcaaaaa cttaatgact ctcaagctcc
     2161 aaaagctgat gcgcaacaaa ataacttcaa caaagatcaa caaagcgcct tctatgaaat
     2221 cttgaacatg cctaacttaa acgaagcgca acgtaacggc ttcattcaaa gtcttaaaga
     2281 cgacccaagc caaagcacta acgttttagg tgaagctaaa aaattaaacg aatctcaagc
     2341 accgaaagct gataacaatt tcaacaaaga acaacaaaat gctttctatg aaatcttgaa
     2401 tatgcctaac ttaaacgaag aacaacgcaa tggtttcatc caaagcttaa aagatgaccc
     2461 aagccaaagt gctaacctat tgtcagaagc taaaaagtta aatgaatctc aagcaccgaa
     2521 agcggataac aaattcaaca aagaacaaca aaatgctttc tatgaaatct tacatttacc
     2581 taacttaaac gaagaacaac gcaatggttt catccaaagc ctaaaagatg acccaagcca
     2641 aagcgctaac cttttagcag aagctaaaaa gctaaatgat gctcaagcac caaaataact
     2701 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
     2761 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     2821 cttgaggggt tttttgctga aaggaggaac tatatccgga tgtggtttgt ccaaactcat
     2881 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2941 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3001 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3061 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3121 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3181 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3241 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3301 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3361 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3421 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3481 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3541 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3601 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3661 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3721 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3781 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4621 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4681 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4741 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4801 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4861 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4921 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4981 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5041 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5101 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5161 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5221 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5281 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5341 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5401 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5461 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5521 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5581 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5641 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5701 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5761 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5821 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5881 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     5941 cattagttca tagcccatat atgg
//
