LOCUS       sequence-399906-        5033 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 5033)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5033)
  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 41kDa (BFP)
FEATURES             Location/Qualifiers
     source          1..5033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        34..337
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        338..541
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     492..512
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      580..589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(628..630,631..1341)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLLE
                     FRTAAGITLGMDELCK"
     primer_bind     complement(670..691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1710..1736
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1749..1766
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1819..1837)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1833..1880
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1906..1925
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1943..2398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2080..2101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2292..2311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2425..2529
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2531..2722
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2554..2574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2719..2853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2902..3696
                     /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             2902..3696
                     /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(2956..2975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3566..3585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3884..3903)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3928..3975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4304..4892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4793..4812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
       61 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      121 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      181 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      241 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      301 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      361 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      421 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      481 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      541 tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggg cccaaaaaag
      601 gccagaaaag ttggaggcgg gggcagcgtg agcaagggcg aggagctgtt caccggggtg
      661 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgaggggc
      721 gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc
      781 aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt gcagtgcttc
      841 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      901 tacgtccagg agcgcaccat cttcttcaag gacgacggca cctacaagac ccgcgccgag
      961 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag
     1021 gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca caacatctat
     1081 atcatggccg tcaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacgtg
     1141 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1201 cccgtgctgc tgcccgacag ccactacctg agcacccagt ccgtgctgag caaagacccc
     1261 aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg gatcactctc
     1321 ggcatggacg agctgtgcaa gaagcttgct caacaaaatg ctttttatca agtcttaaat
     1381 atgcctaact taaatgctga tcaacgcaat ggttttatcc aaagccttaa agatgatcca
     1441 agccaaagtg ctaacgtttt aggtgaagct caaaaactta atgactctca agctccaaaa
     1501 gctgatgcgc aacaaaataa cttcaacaaa gatcaacaaa gcgccttcta tgaaatcttg
     1561 aacatgccta acttaaacga agcgcaacgt aacggcttca ttcaaagtct taaagacgac
     1621 ccaagccaaa gcactaacgt tttaggtgaa gctaaaaaat taaacgaatc tcaagcaccg
     1681 aaagctgata acaattaagt cgacccgggt atccgtatga tgtgcctgac tacgcatgat
     1741 atctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1801 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1861 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatgtggt ttgtccaaac
     1921 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     1981 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2041 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2101 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2161 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2221 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2281 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2341 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2401 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2461 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2521 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2581 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2641 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     2881 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2941 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3001 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3061 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3121 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3181 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3241 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3301 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3361 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3421 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3481 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3541 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3601 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3661 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3721 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     3781 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3841 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     3901 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     3961 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4021 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4081 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4141 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4201 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4261 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4321 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4381 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4441 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4501 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4561 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4621 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4681 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4741 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4801 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4861 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4921 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4981 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggg
//
