LOCUS       sequence-401184-        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Moclo level 1 - Module D, Promoter: CmYLCV, Gene: 
            NbGP41P-sfGFP-VP64-GB1, Terminator: ZmUBI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  AUTHORS   Casas-Mollano JA, Zinselmeier MH, Sychla A, Smanski MJ
  TITLE     Efficient gene activation in plants by the MoonTag programmable 
            transcriptional activator.
  JOURNAL   Nucleic Acids Res. 2023 Jul 21;51(13):7083-7093. doi: 
            10.1093/nar/gkad458.
  PUBMED    37254802
REFERENCE   2  (bases 1 to 5083)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2023-07-21- 21"; volume: "51"; issue: 
            "13"; pages: "7083-7093"
COMMENT     Sequence Label: pMOD_D-CmYLCV-NbGP41P-sfGFP-VP64-GB1
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        67..171
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1239..1827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1728..1747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1981..1998
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             2904..3614
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             2904..3614
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2931..2959)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3554..3574
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3654..3803
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             3828..3848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3879..4040
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             4068..4118
                     /codon_start=1
                     /product="nuclear localization signal from the human T-cell
                     leukemia virus type 1 (HTLV-1) Rex protein"
                     /label=Rex NLS
                     /translation="PKTRRRPRRSQRKRPPT"
ORIGIN
        1 gagctcagtg ttttgcaggt gtgatgaaaa gccttagggt caggtggcac ttttctggga
       61 aatgtgcgat gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgcta
      121 atgagacaat aaccctgata aatgcttcaa ttatattgaa aaaggaagag tatgagtatc
      181 caacatttcc gtgtcgccct tattcacttt tttgcggcct tctgccttcc tgtttttgct
      241 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc ccgagtgggt
      301 tacattgaac tggatctcaa cagcggcaaa atcttagaga gttttcgccc cgaagaacgc
      361 ttcccaatga tgagcacttt caaagttctg ctatgtggcg cggtgttatc ccgtattgat
      421 gctgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
      481 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgtagtgct
      541 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
      601 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      661 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      721 atggcaacaa ctctccgcaa actattaact ggcgaactac ttactcttgc ttcacgccag
      781 caactcattg actggatgga ggcggacaaa gttgcaggac cacttctgcg ctcggcactt
      841 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggctc tcgcggtatc
      901 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      961 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1021 aagcattggt aactgtcaga cgaagggaaa taaatagtag cccgcctgat gtgcgggctt
     1081 ttttttggtg gtggttacca gtgtcgctta gctaccgtag gtcctcttag gcggtccgat
     1141 tagcgacata gtcatcaatg ccttgacggc aaaatccctt aacgtgagtt ttcgttccac
     1201 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1261 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1321 caagagctac taactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1381 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1441 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1501 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1561 gggggttcgt gctcacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1621 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1681 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1741 tatctttata gtcctgtcgg gtttcgccac ctatgacttg agcgtcgatt tttgtgatgc
     1801 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1861 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1921 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1981 agcgagtcag tgagcgagga agcggaagag cgcccacctg ctttttcccg gcgcgccgga
     2041 gtggcagaca tactgtccca caaatgaaga tggaatctgt aaaagaaaac gcgtgaaata
     2101 atgcgtctga caaaggttag gtcggctgcc tttaatcaat accaaagtgg tccctaccac
     2161 gatggaaaaa ctgtgcagtc ggtttggctt tttctgacga acaaataaga ttcgtggccg
     2221 acaggtgggg gtccaccatg tgaaggcatc ttcagactcc aataatggag caatgacgta
     2281 agggcttacg aaataagtaa gggtagtttg ggaaatgtcc actcacccgt cagtctataa
     2341 atacttagcc cctccctcat tgttaaggga gcaaaatctc agagagatag tcctagagag
     2401 agaaagagag caagtagcct agaagtagtc aaggcggcga agtattcagg cacgtggcca
     2461 ggaagaagaa aagccaagac gacgaaaaca ggtaagagct aagcttaatg gaggtgcagc
     2521 tggtggaatc tgggggcgga ctggtgcagc ccgggggatc tctgcggctg tcctgcgccg
     2581 cctctggctc catctctagc gtggatgtga tgtcctggta caggcaggcc cctggcaagc
     2641 agagggaact ggtggccttc attactgata ggggaagaac caattacaaa gtgagcgtga
     2701 aaggccgctt cactatcagc cgggataatt ctaagaatat ggtgtatctg cagatgaaca
     2761 gcctgaagcc agaagacact gccgactatc tgtgcagggc tgagtcccgg acttcctggt
     2821 ccagcccatc tcctctggat gtgtgggggc gcggcactca ggtgactgtg tcctccttgg
     2881 atccaggtgg aggtggaagc ggtagcaaag gagaagaact tttcactgga gttgtcccaa
     2941 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg
     3001 aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact ggaaaactac
     3061 ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc ttttcccgtt
     3121 atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac
     3181 aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct gaagtcaagt
     3241 ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt aaagaagatg
     3301 gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta tacatcacgg
     3361 cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac gttgaagatg
     3421 gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     3481 ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa
     3541 agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca catggcatgg
     3601 atgagctcta caaaggtgga ggtcggaccg gtcgtacggg cggtggcgga ggggatgctt
     3661 tagacgattt tgacttagat atgcttggtt cagacgcgtt agacgacttc gacctagaca
     3721 tgttaggctc agatgcattg gacgacttcg atttagatat gttgggctcc gatgccctag
     3781 atgactttga tctagatatg ctaggtagtg gcggtggcgg aggggctcca aagaagaaga
     3841 gaaaggtccg tacgggagga ggatctcgga ccgaagagta caagcttatc ctgaacggta
     3901 aaaccctgaa aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt
     3961 tcaaacagta cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca
     4021 aaaccttcac ggtaaccgaa ggtggtggta gcggtggtgg tactagtcca aaaacaagga
     4081 ggaggccgcg aagatcacaa cggaaaaggc cgcctacgcc atggccgtaa gcttgtcatg
     4141 ggtcgtttaa gctgccgatg tgcctgcgtc gtctggtgcc ctctctccat atggaggttg
     4201 tcaaagtatc tgctgttcgt gtcatgagtc gtgtcagtgt tggtttaata atggaccggt
     4261 tgtgttgtgt gtgcgtacta cccagaacta tgacaaatca tgaataagtt tgatgtttga
     4321 aattaaagcc tgtgctcatt atgttctgtc tttcagttgt ctcctaatat ttgcctgcag
     4381 gtactggcta tctaccgttt cttacttagg aggtgtttga atgcactaaa actaatagtt
     4441 agtggctaaa attagttaaa acatccaaac accatagcta atagttgaac tattagctat
     4501 ttttggaaaa ttagttaata gtgaggtagt tatttgttag ctagctaatt caactaacaa
     4561 tttttagcca actaacaatt agtttcagtg cattcaaaca cccccttaat gttaacgtgg
     4621 ttctatctac cgtgtcctaa tatatggttg attgttcggt ttgttgctat gctattgggt
     4681 tctgattgct gctagttctt gctgaatcca gaagttctcg tagtatagct cagattcata
     4741 ttatttattt gagtgataag tgatccaggt tattactatg ttagctaggt tttttttaca
     4801 aggataaatt atctgtgatc ataattctta tgaaagcttt atgtttcctg gaggcagtgg
     4861 catgcaatgc atgacagcaa cttgatcaca ccagctgagg tagatacggt aacaaggttc
     4921 ttaaatctgt tcaccaaatc attggagaac acacatacac attcttgcca gtcttggtta
     4981 gagaaatttc atgacaaaat gccaaagctg tcttgactct tcacttttgg ccatgagtcg
     5041 tgacttagtt tggtttaatg gaccggttct cctagcttgc gct
//
