LOCUS       sequence-457132-        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes NLS/cMyc-AcGFP2x-NES/iKB2 which is a tandem of AcGFP with an
            N-terminal cMyc NLS and a C-terminal iKB2 NES that shuttles between 
            the nucleus and the cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  AUTHORS   Vercruysse T, Vanstreels E, Jacquemyn M, Boland S, Kilonda A, 
            Allasia S, Vandecaetsbeek I, Klaassen H, Versele M, Chaltin P, 
            Marchand A, Daelemans D
  TITLE     Ibetazol, a novel inhibitor of importin beta1-mediated nuclear 
            import.
  JOURNAL   Commun Biol. 2024 Nov 23;7(1):1560. doi: 10.1038/s42003-024-07237-8.
  PUBMED    39580542
REFERENCE   2  (bases 1 to 5444)
  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: "Commun 
            Biol."; date: "2024-11-23"; volume: "7(1):1560. doi"; pages: " 
            10.1038/s42003-024-07237-8"
COMMENT     Sequence Label: pGL4-CMV-NLS/cMyc-AcGFP2x-NES/iKB2-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..5444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        221..600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        601..804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     755..775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     849..868
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        849..867
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             912..938
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             948..1661
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     CDS             1680..2396
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     CDS             2472..2528
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    3152..3273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3163..3182)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3217..3236
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3521..3538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3692..4280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3772..3791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4480..5271,5272..5340))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctcttaatga tagctagcgg gccagtaccc
      181 aaaaagcggg gggtctggtg gcagtggagg gggatccgtt gacattgatt attgactagt
      241 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      301 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      361 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      421 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      481 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      541 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      601 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      661 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      721 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      781 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      841 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
      901 ttggtaccat gcctgctgcc aagagggtca agttggacgc tcgaggagtg agcaagggcg
      961 ccgagctgtt caccggcatc gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc
     1021 acaagttcag cgtgagcggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1081 agttcatctg caccaccggc aagctgcctg tgccctggcc caccctggtg accaccctga
     1141 gctacggcgt gcagtgcttc tcacgctacc ccgatcacat gaagcagcac gacttcttca
     1201 agagcgccat gcctgagggc tacatccagg agcgcaccat cttcttcgag gatgacggca
     1261 actacaagtc gcgcgccgag gtgaagttcg agggcgatac cctggtgaat cgcatcgagc
     1321 tgaccggcac cgatttcaag gaggatggca acatcctggg caataagatg gagtacaact
     1381 acaacgccca caatgtgtac atcatgaccg acaaggccaa gaatggcatc aaggtgaact
     1441 tcaagatccg ccacaacatc gaggatggca gcgtgcagct ggccgaccac taccagcaga
     1501 atacccccat cggcgatggc cctgtgctgc tgcccgataa ccactacctg tccacccaga
     1561 gcgccctgtc caaggacccc aacgagaagc gcgatcacat gatctacttc ggcttcgtga
     1621 ccgccgccgc catcacccac ggcatggatg agctgtacaa gaagcttggc gccggtgcaa
     1681 tggtgagcaa gggcgccgag ctgttcaccg gcatcgtgcc catcctgatc gagctgaatg
     1741 gcgatgtgaa tggccacaag ttcagcgtga gcggcgaggg cgagggcgat gccacctacg
     1801 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcctgtgccc tggcccaccc
     1861 tggtgaccac cctgagctac ggcgtgcagt gcttctcacg ctaccccgat cacatgaagc
     1921 agcacgactt cttcaagagc gccatgcctg agggctacat ccaggagcgc accatcttct
     1981 tcgaggatga cggcaactac aagtcgcgcg ccgaggtgaa gttcgagggc gataccctgg
     2041 tgaatcgcat cgagctgacc ggcaccgatt tcaaggagga tggcaacatc ctgggcaata
     2101 agatggagta caactacaac gcccacaatg tgtacatcat gaccgacaag gccaagaatg
     2161 gcatcaaggt gaacttcaag atccgccaca acatcgagga tggcagcgtg cagctggccg
     2221 accactacca gcagaatacc cccatcggcg atggccctgt gctgctgccc gataaccact
     2281 acctgtccac ccagagcgcc ctgtccaagg accccaacga gaagcgcgat cacatgatct
     2341 acttcggctt cgtgaccgcc gccgccatca cccacggcat ggatgagctg tacaagggag
     2401 ccggagctcc aagcacccgg atacagcagc agctgggcca gctgacacta gaaaaccttc
     2461 agggtagtgg tgccacgaat tttagtctcc tcaagcaagc tggggacgtc gaggagaatc
     2521 ctggcccaat gactgaatac aagcctactg tcaggttggc tacaagagac gacgttccta
     2581 gagccgtgag aactctggct gcagccttcg ccgactaccc cgccacgaga cacaccgttg
     2641 acccagatcg gcatattgag agagtgactg aactgcagga gctgtttctt acaagagttg
     2701 gcctcgacat aggcaaggtg tgggtggcgg acgacggcgc cgccgtggcc gtctggacca
     2761 ctcccgaatc agttgaggct ggcgccgtat tcgctgagat cggcccgaga atggctgagc
     2821 tcagcgggag taggctcgcg gcacagcagc aaatggaggg actgctggca ccacacaggc
     2881 ccaaagaacc cgcctggttc ctggcaaccg tcggtgtatc tcccgatcat caggggaaag
     2941 gtctgggctc tgccgtagtg ctccctggcg tggaggcagc tgagagagca ggagtacctg
     3001 ccttcttgga gacctccgct ccaaggaatc ttcccttcta tgaacggttg ggcttcaccg
     3061 tgacagccga cgtggaagtc cccgaaggcc cccgcacttg gtgcatgacg aggaagcctg
     3121 gagcgtgagg gcccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3181 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3241 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3301 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3361 ggtaaaatcg ataaagatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     3421 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     3481 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     3541 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3601 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3661 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3721 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3781 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3841 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3901 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3961 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4021 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4081 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4141 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4201 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4261 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4321 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4381 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4441 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     4501 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     4561 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     4621 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     4681 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     4741 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     4801 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     4861 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     4921 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     4981 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     5041 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     5101 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     5161 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     5221 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     5281 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     5341 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
     5401 ataagtaagt aatattaagg tacgggaggt attggacagg ccgc
//
