LOCUS       sequence-128350-        4858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for Yellow Flurescent Protein plant expression 
            driven by the 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4858)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 4858)
  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 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: pEGB 35S:YFP:Tnos (GB0209)
FEATURES             Location/Qualifiers
     source          1..4858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(1033..1035,1036..1038,1039..1749)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1033..1035,1036..1038,1039..1749)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1078..1099)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1339..1358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1686..1707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1788..2040
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2075..2093)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2281..2305
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2396..2984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2476..2495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3155..3970)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3878..3897
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3962..3981
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4261..4696
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4823..4845
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat accacaacaa aacaaacaac aaacaacatt acaatttact
     1021 attctagtcg aaatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1081 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1141 gacgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1201 ccctggccca ccctcgtgac caccttcggc tacggcctga agtgcttcgc ccgctacccc
     1261 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1321 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1381 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1441 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1501 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1561 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1621 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1681 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1741 ctctacaagt cagtcatgag ctgagcttgg aatggatctt cgatcccgat cgttcaaaca
     1801 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgacg attatcatat
     1861 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     1921 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     1981 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     2041 gggaattgcc aagctaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     2101 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2161 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2221 caataaccct gataaatgct tcaataatgg gaccgactcg cgctgtcatg agacgaattc
     2281 tgacaggata tattggcggg taaactaagt cgctgtatgt gtttgtttga gatctcatgt
     2341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2881 gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3121 tctaaagtat atatgtgtaa cattggtcta gtgattagaa aaactcatcg agcatcaaat
     3181 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     3241 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     3301 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     3361 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
     3421 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     3481 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagtcga aatacgcgat
     3541 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     3601 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     3661 tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     3721 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacaa
     3781 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     3841 acaatcggta gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     3901 ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt tcccgttgaa
     3961 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     4021 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
     4081 gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg
     4141 cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag
     4201 ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcgatcc
     4261 ccatccaaca gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt ggatagaggg
     4321 tagttatcca cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg gctttccggc
     4381 ccgctccaaa aactatccac gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga
     4441 gtacgtgaaa tcgctaataa ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg
     4501 ctaatagccc tttcagatca acagcttgca aacacccctc gctccggcaa gtagttacag
     4561 caagtagtat gttcaattag cttttcaatt atgaatatat atatcaatta ttggtcgccc
     4621 ttggcttgtg gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt
     4681 gcccaccgtc gagcgccttt gcccacaacc cggcggccgg ccgcaacaga tcgttttata
     4741 aatttttttt tttgaaaaag aaaaagcccg aaaggcggca acctctcggg cttctggatt
     4801 tccgatcccc ggaattagat cttggcagga tatattgtgg tgtaacgaat tcgtctca
//
