LOCUS       sequence-269204-        4159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for Lifeact-Citrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4159)
  AUTHORS   Kimura S, Kodama Y
  TITLE     Actin-dependence of the chloroplast cold positioning response in the
            liverwort Marchantia polymorpha L.
  JOURNAL   PeerJ. 2016 Sep 28;4:e2513. eCollection 2016.
  PUBMED    27703856
REFERENCE   2  (bases 1 to 4159)
  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: "PeerJ."; 
            date: "2016-09-28"; volume: "4"; pages: "e2513. eCollection 2016"
COMMENT     Sequence Label: pDONR207-Lifeact-Citrine
FEATURES             Location/Qualifiers
     source          1..4159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    31..130
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             133..183
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             join(199..201,202..204,205..915)
                     /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(244..265)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(505..524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     852..873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(920..1019)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1131..1150)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1209..1228)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1802..2335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2524..2529,2530..2546,2547..2552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3209..3797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3698..3717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3932..3959)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4051..4137)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgcgttaacg ctagcatgga tctcgggccc caaataatga ttttattttg actgatagtg
       61 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
      121 aaagcaggct tcatgggcgt ggccgacctg atcaagaagt tcgagagcat cagcaaggag
      181 gagggtggct ctggaggtat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      241 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      301 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      361 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc
      421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      481 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      601 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      661 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      721 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      781 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
      841 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      901 gacgagctgt acaagtgaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
      961 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1021 catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1081 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1141 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1201 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1261 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1321 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1381 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1441 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1501 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1561 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1621 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1681 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     1741 taaccttatt tttgacgagg ggaaattagg ccgggaagcc gatctcggct tgaacgaatt
     1801 gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact
     1861 ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag
     1921 caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc
     1981 tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa
     2041 acttgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa
     2101 gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt
     2161 gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg
     2221 atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta
     2281 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg
     2341 ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga
     2401 ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     2461 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
     2521 tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac
     2581 ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg
     2641 gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc
     2701 ttctacggca aggtgctgtg cacggatctg ccctggcttc aggagatcgg aagacctcgg
     2761 ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt
     2821 ctggaaggcg agcatcgttt gttcgcccag gactctagct atagttctag tggttggcta
     2881 ctaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2941 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3001 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3061 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     3121 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3181 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3241 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3301 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3361 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3421 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3481 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3541 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3601 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3661 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3721 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3781 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3841 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3901 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     3961 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     4021 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     4081 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
     4141 cctggcagtt ccctactct
//
