LOCUS       sequence-181839-        5233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module A, Promoter: 35S, Gene: TALE-VP64_1 backbone with Esp3I ccdb 
            cassette for repeat cloning, Terminator: none (ends with P2A to be 
            fused to TALE-VP64_ 2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5233)
  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 5233)
  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_A3301
FEATURES             Location/Qualifiers
     source          1..5233
                     /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"
     promoter        1366..1711
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1654..1674
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1745..1810
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1817..1837
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2585..2890)
                     /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(2687..2706)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3874..4023
                     /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"
     promoter        4141..4245
                     /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 gctcatgatc ggcgcgccag atttgccttt tcaatttcag
      901 aaagaatgct aacccacaga tggttagaga ggcttacgca gcaggtatca tcaagacgat
      961 ctacccgagc aataatctcc aggaaatcaa ataccttccc aagaaggtta aagatgcagt
     1021 caaaagattc aggactaact gcatcaagaa cacagagaaa gatatatttc tcaagatcag
     1081 aagtactatt ccagtatgga cgattcaagg cttgcttcac aaaccaaggc aagtaataga
     1141 gattggagtc tctaaaaagg tagttcccac tgaatcaaag gccatggagt caaagattca
     1201 aatagaggac ctaacagaac tcgccgtaaa gactggcgaa cagttcatac agagtctctt
     1261 acgactcaat gacaagaaga aaatcttcgt caacatggtg gagcacgaca cacttgtcta
     1321 ctccaaaaat atcaaagata cagtctcaga agaccaaagg gcaattgaga cttttcaaca
     1381 aagggtaata tccggaaacc tcctcggatt ccattgccca gctatctgtc actttattgt
     1441 gaagatagtg gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc
     1501 catcgttgaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
     1561 catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat
     1621 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     1681 ataaggaagt tcatttcatt tggagagaac acgggggact cctgcaggaa aaatggcttc
     1741 ctccgattat aaggatcacg atggcgacta caaggaccac gatattgact acaaagacga
     1801 tgacgataaa atggctccta agaaaaagcg caaagtcggt atccatggcg ttcccgctag
     1861 cgtgtcccgg acccggctgc catctccccc tgcgccctct cctgcgttct cggcgggcag
     1921 cttcagcgat ctgctccgtc cgttcgatcc gtcgcttctt gatacatcgc ttcttgattc
     1981 gatgcctgcc gtcggcacgc cgcatacagc ggctgcccca gcagagtggg atgaggcgca
     2041 atcggctctg cgtgcagccg atgacccgcc acccaccgtg cgtgtcgctg tcactgccgc
     2101 gcggccgccg cgcgccaagc cggccccgcg acggcgtgct gcgcaaccct ccgacgcttc
     2161 gccggccgcg caggttgatc tacgcacgct cggctacagt cagcagcagc aagagaagat
     2221 caaaccgaag gtgcgttcga cagtggcgca gcaccacgag gcactggtgg gccatgggtt
     2281 tacacacgcg cacatcgttg cgctcagcca acacccggca gcgttaggga ccgtcgctgt
     2341 cacgtatcag cacataatca cggcgttgcc agaggcgaca cacgaagaca tcgttggcgt
     2401 cggcaaacag tggtccggcg cacgcgccct ggaggccttg ctcacggatg cgggggagtt
     2461 gagaggtccg ccgttacagt tggacacagg ccaacttgtg aagattgcaa aacgtggcgg
     2521 cgtgaccgca atggaggcag tgcatgccag caggaatgcg ttaacaggtg cccccctgga
     2581 gacgttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc
     2641 tgagatcagc cacttcttcc ccgataacgg aaaccggcac actggccata tcggtggtca
     2701 tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg gagactttat
     2761 ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc gtgtcaataa
     2821 tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag gtgtaaacct
     2881 taaactgcat ttcaccagcc cctgttctcg tcagcaaaag agccgttcat ttcaataaac
     2941 cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc acgcagacga
     3001 cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc atatatgcct
     3061 tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc atagcatacc
     3121 tccttttgac atacttcggg tatacatatc agtatatatt cttataccgc aaaaatcagc
     3181 gcgcaaatac gcatactgtt atctggcttc gtctccaacg accacctcgt cgccttggcc
     3241 tgcctcggcg gacgtcctgc catggatgca gtgaaaaagg gattgccgca cgcgccggaa
     3301 ttgatcagaa gagtcaatcg ccgtattggc gaacgcacgt cccatcgcgt tgccgactac
     3361 gcgcaagtgg ttcgcgtgct ggagtttttc cagtgccact cccacccagc gtacgcattt
     3421 gatgaggcca tgacgcagtt cgggatgagc aggaacgggt tggtacagct ctttcgcaga
     3481 gtgggcgtca ccgaactcga agcccgcggt ggaacgctcc ccccagcctc gcagcgttgg
     3541 gaccgtatcc tccaggcatc agggatgaaa agggccaaac cgtcccctac ttcagctcaa
     3601 acaccggatc aggcgtcttt gcatgcattc gccgattcgc tggagcgtga ccttgatgcg
     3661 cccagcccaa tgcacgaggg agatcagacg cgggcaagca gccgtaaacg gtcccgatcg
     3721 gatcgtgctg tcaccggccc ctccgcacag caggctgtcg aggtgcgcgt tcccgaacag
     3781 cgcgatgcgc tgcatttgcc cctcagctgg agggtaaaac gcccgcgtac caggatctgg
     3841 ggcggcctcc cggatccggg ttcaggacgg gctgacgcat tggacgattt tgatctggat
     3901 atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt
     3961 gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg
     4021 ctgattaact ctagaagttc cggaagcggc gccacgaact tctctctgtt aaagcaagca
     4081 ggggacgtgg gcaggtgatg aaaagcctta gggtcaggtg gcacttttcg gggaaatgtg
     4141 cgatgaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctaatgaga
     4201 caataaccct gataaatgct tcaattatat tgaaaaagga agagtatgag tatccaacat
     4261 ttccgtgtcg cccttattcc cttttttgcg gccttctgcc ttcctgtttt tgctcaccca
     4321 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcccgagt gggttacatt
     4381 gaactggatc tcaacagcgg caaaatctta gagagttttc gccccgaaga acgcttccca
     4441 atgatgagca ctttcaaagt tctgctatgt ggcgcggtgt tatcccgtat tgatgctggg
     4501 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     4561 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgtag tgctgccata
     4621 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     4681 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4741 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4801 acaactctcc gcaaactatt aactggcgaa ctacttactc ttgcttcacg ccagcaactc
     4861 attgactgga tggaggcgga caaagttgca ggaccacttc tgcgctcggc acttccggct
     4921 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4981 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5041 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5101 tggtaactgt cagacgaagg gaaataaata gtagcccgcc tgatgtgcgg gctttttttt
     5161 ggtggtggtt accagtgtcg cttagctacc gtaggtcctc ttaggcggtc cgattagcga
     5221 catagtcatc aat
//
