LOCUS       sequence-203339-        4767 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes half of a protein delivery sensor ( NLS-FKBP-VN-T2A-RFP) for
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4767)
  AUTHORS   Chiu HY, Bates JA, Helma J, Engelke H, Harz H, Bein T, Leonhardt H
  TITLE     Nanoparticle Mediated Delivery and Small Molecule Triggered 
            Activation of Proteins in the Nucleus.
  JOURNAL   Nucleus. 2018 Sep 14. doi: 10.1080/19491034.2018.1523665.
  PUBMED    30217128
REFERENCE   2  (bases 1 to 4767)
  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"; journalName: "Nucleus. 
            2018 Sep 14. doi: 10.1080/19491034.2018.1523665."
COMMENT     Sequence Label: pCAG-c-Myc NLS-FKBP-Venus N-T2A-mRFP
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    91..553
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(147..164)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     374..393
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             564..962
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1120..1241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1157..1176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1211..1230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1289..1424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1530..1547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1701..2289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1781..1800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2460..3251,3252..3320))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3083..3102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3321..3425)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3493..3511
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3549..3571)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3671..3690
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3757..4212)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3894..3915)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4106..4125
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4338..4360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4352..4369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4353..4369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4376..4394
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        4463..4766
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
       61 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
      121 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
      181 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
      241 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
      301 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
      361 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
      421 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
      481 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
      541 aacacgatga taagcttgcc accatggcca agcctttgtc tcaagaagaa tccaccctca
      601 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
      661 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
      721 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
      781 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
      841 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
      901 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
      961 aagcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     1021 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     1081 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     1141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     1261 acctcgagga gctttttgca aaagcctagg cctccaaaaa agcctcttca ctacttctgg
     1321 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     1381 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     1441 gggactatgg ttgctgacta attgctgcat taatgaatcg gccaacgcgc ggggagaggc
     1501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2581 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3421 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3481 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     3541 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3601 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     3661 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     3721 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     3781 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3841 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3901 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3961 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4021 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4081 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4141 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctc aatgcgccgc
     4201 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4261 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4321 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata
     4381 cgactcacta taggacgcgt cctagttatt aatagtaatc aattacgggg tcattagttc
     4441 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4501 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4561 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4621 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4681 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4741 acgtattagt catcgctatt accatgg
//
