LOCUS       sequence-399879-        5054 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 5054)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5054)
  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_NLS 41kDa (BFP)
FEATURES             Location/Qualifiers
     source          1..5054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        39..342
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        343..546
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     497..517
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      585..594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             597..617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(633..635,636..1346)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="VCKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLLE
                     FRTAAGITLGMDELYK"
     primer_bind     complement(675..696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1715..1741
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1754..1771
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1824..1842)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1838..1885
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1911..1930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1948..2403)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2085..2106)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2297..2316
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2430..2534
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2536..2893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2536..2727
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2559..2579)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2744..2879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2928..3722
                     /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             2928..3722
                     /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(2982..3001)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3592..3611
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3910..3929)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3954..4001
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4330..4918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4819..4838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
       61 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      121 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      181 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      241 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      301 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      361 tacatcattg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      421 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      481 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      541 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atgggcccaa
      601 aaaagaaaag aaaagttgga ggcgggggca gcgtgtgcaa gggcgaggag ctgttcaccg
      661 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtga
      721 ggggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
      781 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgagccac ggcgtgcagt
      841 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      901 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg
      961 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcgtcgact
     1021 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaaca
     1081 tctatatcat ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1141 acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1201 acggccccgt gctgctgccc gacagccact acctgagcac ccagtccgtg ctgagcaaag
     1261 accccaacga gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc gccgggatca
     1321 ctctcggcat ggacgagctg tacaagaagc ttgctcaaca aaatgctttt tatcaagtct
     1381 taaatatgcc taacttaaat gctgatcaac gcaatggttt tatccaaagc cttaaagatg
     1441 atccaagcca aagtgctaac gttttaggtg aagctcaaaa acttaatgac tctcaagctc
     1501 caaaagctga tgcgcaacaa aataacttca acaaagatca acaaagcgcc ttctatgaaa
     1561 tcttgaacat gcctaactta aacgaagcgc aacgtaacgg cttcattcaa agtcttaaag
     1621 acgacccaag ccaaagcact aacgttttag gtgaagctaa aaaattaaac gaatctcaag
     1681 caccgaaagc tgataacaat taagtcgacc cgggtatccg tatgatgtgc ctgactacgc
     1741 atgatatctc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     1801 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1861 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat gtggtttgtc
     1921 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     1981 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2041 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2101 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2161 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2221 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2281 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2341 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2401 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2461 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2521 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     2581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2641 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2821 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2881 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2941 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3001 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3061 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3121 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3181 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3241 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3301 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3361 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3421 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     3481 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3541 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3601 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3661 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3721 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     3781 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3841 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     3901 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3961 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4021 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4081 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4141 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4201 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4261 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4321 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4381 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4441 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4501 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4561 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4621 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4681 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4741 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4801 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4861 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4921 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4981 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     5041 aatagtaatc aatt
//
