LOCUS       sequence-182042-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: none (begins with P2A to be fused to 
            TALE-VPR_1), Gene: TALE-VPR_2 backbone with Esp3I ccdb cassette for 
            repeat cloning, Terminator: HSP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 6011)
  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.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pMOD_B3200
FEATURES             Location/Qualifiers
     source          1..6011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             895..960
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             967..987
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1735..2040)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1837..1856)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3024..3173
                     /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             3198..3218
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3648..4004
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             4023..4592
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     terminator      4607..4855
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        4919..5023
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcggaaga gcgcccacct gccaggggac gtggaagaaa acccagggcc catggactac
      901 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
      961 atggccccca agaagaagag gaaggtgggc attcacgggg tgccggctag cgtgtcccgg
     1021 acccggctgc catctccccc tgcgccctct cctgcgttct cggcgggcag cttcagcgat
     1081 ctgctccgtc cgttcgatcc gtcgcttctt gatacatcgc ttcttgattc gatgcctgcc
     1141 gtcggcacgc cgcatacagc ggctgcccca gcagagtggg atgaggcgca atcggctctg
     1201 cgtgcagccg atgacccgcc acccaccgtg cgtgtcgctg tcactgccgc gcggccgccg
     1261 cgcgccaagc cggccccgcg acggcgtgct gcgcaaccct ccgacgcttc gccggccgcg
     1321 caggttgatc tacgcacgct cggctacagt cagcagcagc aagagaagat caaaccgaag
     1381 gtgcgttcga cagtggcgca gcaccacgag gcactggtgg gccatgggtt tacacacgcg
     1441 cacatcgttg cgctcagcca acacccggca gcgttaggga ccgtcgctgt cacgtatcag
     1501 cacataatca cggcgttgcc agaggcgaca cacgaagaca tcgttggcgt cggcaaacag
     1561 tggtccggcg cacgcgccct ggaggccttg ctcacggatg cgggggagtt gagaggtccg
     1621 ccgttacagt tggacacagg ccaacttgtg aagattgcaa aacgtggcgg cgtgaccgca
     1681 atggaggcag tgcatgcatc gcgcaatgca ctgacgggtg cccccctgga gacgttatat
     1741 tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc tgagatcagc
     1801 cacttcttcc ccgataacgg aaaccggcac actggccata tcggtggtca tcatgcgcca
     1861 gctttcatcc ccgatatgca ccaccgggta aagttcacgg gagactttat ctgacagcag
     1921 acgtgcactg gccaggggga tcaccatccg tcgcccgggc gtgtcaataa tatcactctg
     1981 tacatccaca aacagacgat aacggctctc tcttttatag gtgtaaacct taaactgcat
     2041 ttcaccagcc cctgttctcg tcagcaaaag agccgttcat ttcaataaac cgggcgacct
     2101 cagccatccc ttcctgattt tccgctttcc agcgttcggc acgcagacga cgggcttcat
     2161 tctgcatggt tgtgcttacc agaccggaga tattgacatc atatatgcct tgagcaactg
     2221 atagctgtcg ctgtcaactg tcactgtaat acgctgcttc atagcatacc tccttttgac
     2281 atacttcggg tatacatatc agtatatatt cttataccgc aaaaatcagc gcgcaaatac
     2341 gcatactgtt atctggcttc gtctccaacg atcatctggt ggcactcgcg tgcctcggcg
     2401 gacgtcctgc catggatgca gtgaaaaagg gattgccgca cgcgccggaa ttgatcagaa
     2461 gagtcaatcg ccgtattggc gaacgcacgt cccatcgcgt tgccgactac gcgcaagtgg
     2521 ttcgcgtgct ggagtttttc cagtgccact cccacccagc gtacgcattt gatgaggcca
     2581 tgacgcagtt cgggatgagc aggaacgggt tggtacagct ctttcgcaga gtgggcgtca
     2641 ccgaactcga agcccgcggt ggaacgctcc ccccagcctc gcagcgttgg gaccgtatcc
     2701 tccaggcatc agggatgaaa agggccaaac cgtcccctac ttcagctcaa acaccggatc
     2761 aggcgtcttt gcatgcattc gccgattcgc tggagcgtga ccttgatgcg cccagcccaa
     2821 tgcacgaggg agatcagacg cgggcaagca gccgtaaacg gtcccgatcg gatcgtgctg
     2881 tcaccggccc ctccgcacag caggctgtcg aggtgcgcgt tcccgaacag cgcgatgcgc
     2941 tgcatttgcc cctcagctgg agggtaaaac gcccgcgtac caggatctgg ggcggcctcc
     3001 cggatccggg ttcaggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa
     3061 gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg
     3121 acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact
     3181 ctagaagttc cggatctccg aaaaagaaac gcaaagttgg tagccagtac ctgcccgaca
     3241 ccgacgaccg gcaccggatc gaggaaaagc ggaagcggac ctacgagaca ttcaagagca
     3301 tcatgaagaa gtcccccttc agcggcccca ccgaccctag acctccacct agaagaatcg
     3361 ccgtgcccag cagatccagc gccagcgtgc caaaacctgc cccccagcct taccccttca
     3421 ccagcagcct gagcaccatc aactacgacg agttccctac catggtgttc cccagcggcc
     3481 agatctctca ggcctctgct ctggctccag cccctcctca ggtgctgcct caggctcctg
     3541 ctcctgcacc agctccagcc atggtgtctg cactggctca ggcaccagca cccgtgcctg
     3601 tgctggctcc tggacctcca caggctgtgg ctccaccagc ccctaaacct acacaggccg
     3661 gcgagggcac actgtctgaa gctctgctgc agctgcagtt cgacgacgag gatctgggag
     3721 ccctgctggg aaacagcacc gatcctgccg tgttcaccga cctggccagc gtggacaaca
     3781 gcgagttcca gcagctgctg aaccagggca tccctgtggc ccctcacacc accgagccca
     3841 tgctgatgga ataccccgag gccatcaccc ggctcgtgac aggcgctcag aggcctcctg
     3901 atccagctcc tgcccctctg ggagcaccag gcctgcctaa tggactgctg tctggcgacg
     3961 aggacttcag ctctatcgcc gacatggact tctccgcact gctgggtagc ggatcgggat
     4021 ctcgggattc cagggaaggg atgtttttgc cgaagcctga ggccggctcc gctattagtg
     4081 acgtgtttga gggccgcgag gtgtgccagc caaaacgaat ccggccattt catcctccag
     4141 gaagtccatg ggccaaccgc ccactccccg ccagcctcgc accaacacca accggtccag
     4201 tacatgagcc agtcgggtca ctgaccccgg caccagtccc tcagccactg gatccagcgc
     4261 ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc cgatgaagag actagccagg
     4321 ctgtcaaagc ccttcgggag atggccgata ctgtgattcc ccagaaggaa gaggctgcaa
     4381 tctgtggcca aatggacctt tcccatccgc ccccaagggg ccatctggat gagctgacaa
     4441 ccacacttga gtccatgacc gaggatctga acctggactc acccctgacc ccggaattga
     4501 acgagattct ggataccttc ctgaacgacg agtgcctctt gcatgccatg catatcagca
     4561 caggactgtc catcttcgac acatctctgt tctagatgac tcgagatatg aagatgaaga
     4621 tgaaatattt ggtgtgtcaa ataaaaagct tgtgtgctta agtttgtgtt tttttcttgg
     4681 cttgttgtgt tatgaatttg tggctttttc taatattaaa tgaatgtaag atcacattat
     4741 aatgaataaa caaatgtttc tataatccat tgtgaatgtt ttgttggatc tcttctgcag
     4801 catataacta ctgtatgtgc tatggtatgg actatggaat atgattaaag ataaggagct
     4861 ccggtgacgc aggtgatgaa aagccttagg gtcaggtggc acttttcggg gaaatgtgcg
     4921 atgaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc taatgagaca
     4981 ataaccctga taaatgcttc aattatattg aaaaaggaag agtatgagta tccaacattt
     5041 ccgtgtcgcc cttattccct tttttgcggc cttctgcctt cctgtttttg ctcacccaga
     5101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcccgagtgg gttacattga
     5161 actggatctc aacagcggca aaatcttaga gagttttcgc cccgaagaac gcttcccaat
     5221 gatgagcact ttcaaagttc tgctatgtgg cgcggtgtta tcccgtattg atgctgggca
     5281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     5341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgtagtg ctgccataac
     5401 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     5461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5581 aactctccgc aaactattaa ctggcgaact acttactctt gcttcacgcc agcaactcat
     5641 tgactggatg gaggcggaca aagttgcagg accacttctg cgctcggcac ttccggctgg
     5701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5881 gtaactgtca gacgaaggga aataaatagt agcccgcctg atgtgcgggc ttttttttgg
     5941 tggtggttac cagtgtcgct tagctaccgt aggtcctctt aggcggtccg attagcgaca
     6001 tagtcatcaa t
//
