LOCUS       sequence-401187-        5306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Moclo level 1 - Module D, Promoter: AtUBI10, Gene: 
            NbGP41P-sfGFP-VP64-GB1, Terminator: pea rbcsE9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5306)
  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 5306)
  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-AtUBI10-NbGP41-sfGFP-VP64-GB1
FEATURES             Location/Qualifiers
     source          1..5306
                     /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             3766..4476
                     /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             3766..4476
                     /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(3793..3821)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4416..4436
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4516..4665
                     /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             4690..4710
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4741..4902
                     /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             4930..4980
                     /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 ggtcgagctg caggtcaacg gatcaggata ttcttgttta agatgttgaa ctctatggag
     2101 gtttgtatga actgatgatc taggaccgga taagttccct tcttcatagc gaacttattc
     2161 aaagaatgtt ttgtgtatca ttcttgttac attgttatta atgaaaaaat attattggtc
     2221 attggactga acacgagtgt taaatatgga ccaggcccca aataagatcc attgatatat
     2281 gaattaaata acaagaataa atcgagtcac caaaccactt gcctttttta acgagacttg
     2341 ttcaccaact tgatacaaaa gtcattatcc tatgcaaatc aataatcata caaaaatatc
     2401 caataacact aaaaaattaa aagaaatgga taatttcaca atatgttata cgataaagaa
     2461 gttacttttc caagaaattc actgatttta taagcccact tgcattagat aaatggcaaa
     2521 aaaaaacaaa aaggaaaaga aataaagcac gaagaattct agaaaatacg aaatacgctt
     2581 caatgcagtg ggacccacgg ttcaattatt gccaattttc agctccaccg tatatttaaa
     2641 aaataaaacg ataatgctaa aaaaatataa atcgtaacga tcgttaaatc tcaacggctg
     2701 gatcttatga cgaccgttag aaattgtggt tgtcgacgag tcagtaataa acggcgtcaa
     2761 agtggttgca gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc
     2821 tcaacctaaa aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac
     2881 acgtgtcatt ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt
     2941 cctcgtcttt tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca
     3001 attcagattt caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc
     3061 aaggtaaatt tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc
     3121 caatttcgta tatgttcttt ggtttagatt ctgttaatct tagatcgaac acgattttct
     3181 gggtttgatc gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat
     3241 ttgttcaaat aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag
     3301 atctggtgtt agtttctagt ttgtgcgatc gaatttgtcg attaatctga gtttttctga
     3361 ttaacaggaa tggaggtgca gctggtggaa tctgggggcg gactggtgca gcccggggga
     3421 tctctgcggc tgtcctgcgc cgcctctggc tccatctcta gcgtggatgt gatgtcctgg
     3481 tacaggcagg cccctggcaa gcagagggaa ctggtggcct tcattactga taggggaaga
     3541 accaattaca aagtgagcgt gaaaggccgc ttcactatca gccgggataa ttctaagaat
     3601 atggtgtatc tgcagatgaa cagcctgaag ccagaagaca ctgccgacta tctgtgcagg
     3661 gctgagtccc ggacttcctg gtccagccca tctcctctgg atgtgtgggg gcgcggcact
     3721 caggtgactg tgtcctcctt ggatccaggt ggaggtggaa gcggtagcaa aggagaagaa
     3781 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     3841 ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt
     3901 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat
     3961 ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt
     4021 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac
     4081 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag
     4141 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac
     4201 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa
     4261 attcgccaca acgttgaaga tggttccgtt caactagcag accattatca acaaaatact
     4321 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc
     4381 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct
     4441 gctgggatta cacatggcat ggatgagctc tacaaaggtg gaggtcggac cggtcgtacg
     4501 ggcggtggcg gaggggatgc tttagacgat tttgacttag atatgcttgg ttcagacgcg
     4561 ttagacgact tcgacctaga catgttaggc tcagatgcat tggacgactt cgatttagat
     4621 atgttgggct ccgatgccct agatgacttt gatctagata tgctaggtag tggcggtggc
     4681 ggaggggctc caaagaagaa gagaaaggtc cgtacgggag gaggatctcg gaccgaagag
     4741 tacaagctta tcctgaacgg taaaaccctg aaaggtgaaa ccaccaccga agctgttgac
     4801 gctgctaccg cggaaaaagt tttcaaacag tacgctaacg acaacggtgt tgacggtgaa
     4861 tggacctacg acgacgctac caaaaccttc acggtaaccg aaggtggtgg tagcggtggt
     4921 ggtactagtc caaaaacaag gaggaggccg cgaagatcac aacggaaaag gccgcctacg
     4981 ccatggccgt aagcttgttc gagtattatg gcattgggaa aactgttttt cttgtaccat
     5041 ttgttgtgct tgtaatttac tgtgtttttt attcggtttt cgctatcgaa ctgtgaaatg
     5101 gaaatggatg gagaagagtt aatgaatgat atggtccttt tgttcattct caaattaata
     5161 ttatttgttt tttctcttat ttgttgtgtg ttgaatttga aattataaga gatatgcaaa
     5221 cattttgttt tgagtaaaaa tgtgtcaaat cgtggcctct aatgaccgaa gttaatatga
     5281 ggagtaaaac acttgtagtt gtcgct
//
