LOCUS       sequence-158537-        4117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for Citrine fluorescent protein with a nuclear 
            localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4117)
  AUTHORS   Ogasawara Y, Ishizaki K, Kohchi T, Kodama Y
  TITLE     Cold-induced organelle relocation in the liverwort Marchantia 
            polymorpha L.
  JOURNAL   Plant Cell Environ. 2013 Feb 20. doi: 10.1111/pce.12085.
  PUBMED    23421791
REFERENCE   2  (bases 1 to 4117)
  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: "Plant Cell 
            Environ. 2013 Feb 20. doi: 10.1111/pce.12085."
COMMENT     Sequence Label: pDONR207-Citrine-NLS
FEATURES             Location/Qualifiers
     source          1..4117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(73..100)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(192..278)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    331..430
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(433..435,436..438,439..1149)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(478..499)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(739..758)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1086..1107
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1153..1173
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1178..1277)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1389..1408)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1467..1486)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2060..2593)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2782..2787,2788..2804,2805..2810))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3467..4055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3956..3975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagcc aggaagagtt
       61 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
      121 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
      181 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      241 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      301 cgcgttaacg ctagcatgga tctcgggccc caaataatga ttttattttg actgatagtg
      361 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaag tttgtacaaa
      421 aaagcaggct ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      481 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      541 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      601 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc
      661 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      721 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      781 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      841 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      901 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      961 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1021 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1081 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1141 ctgtacaagc ctcctaagaa gaagcgcaag gtgtgagacc cagctttctt gtacaaagtt
     1201 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1261 aataaaatca ttatttgcca tccagctgca gctctggccc gtgtctcaaa atctctgatg
     1321 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1381 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1441 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1501 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1561 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1621 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1681 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1741 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1801 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1861 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1921 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1981 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaggcc gggaagccga
     2041 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
     2101 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
     2161 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
     2221 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
     2281 cactacgcgg ctgctcaaac ctgggcagaa cgtaagccgc gagagcgcca acaaccgctt
     2341 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
     2401 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
     2461 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
     2521 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     2581 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
     2641 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
     2701 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     2761 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt
     2821 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag
     2881 gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca
     2941 ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc ctggcttcag
     3001 gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg
     3061 gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat
     3121 agttctagtg gttggctact aataggttgt attgatgttg gacgagtcgg aatcgcagac
     3181 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     3241 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3301 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3361 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagtttt
     3421 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3481 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3541 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3601 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3661 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3721 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3781 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3841 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3901 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3961 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4021 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4081 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
