LOCUS       sequence-181878-        5093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: PvUbi1, Gene: SapI ccdb cassette for cloning 
            multiple gRNA spacers with tRNA spacers , Terminator: 35S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5093)
  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 5093)
  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_B2312
FEATURES             Location/Qualifiers
     source          1..5093
                     /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             complement(1589..1600)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        2945..3020
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             3417..3722
                     /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     3601..3620
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    3750..3924
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        4001..4105
                     /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 aagcgggaga gcgcccacct gccaggggac ggcgcgccga agagcacgtc agtgtttggt
      901 ttccactagc acgagtagcg caatcagaaa attttcaatg catgaagtac taaacgaagt
      961 ttatttagaa atttttttaa gaaatgagtg taattttttg cgacgaattt aatgacaata
     1021 attaatcgat gattgcctac agtaatgcta cagtaaccaa cctctaatca tgcgtcgaat
     1081 gcgtcattag attcgtctcg caaaatagca caagaattat gaaattaatt ttacaaacta
     1141 tttttattta atactaataa ttaactgtca aagtttgtgc tactcgcaag agtagcgcga
     1201 accaaacacg gcctggagga gcacggtaac ggcgtcgaca aactaacggc caccacccgc
     1261 caacgcaaag gagacggatg agagttgact tcttgacggt tctccacccc tctgtctctc
     1321 tgtcactggg ccctgggtcc ccctctcgaa agttcctctg gccgaaattg cgcggcggag
     1381 acgaggcggg cggaaccgtc acggcagagg attccttccc caccctgcct ggcccggcca
     1441 tatataaaca gccaccgccc ctccccgttc cccatcgcgt ctcgtctcgt gttgttccca
     1501 gaacacaacc aaaatccaaa tcctcctcct cctcccgagc ctcgtcgatc cctcacccgc
     1561 ttcaaggtac ggcgatcctc ctctcccttc tcccctcgat cgattatgcg tgttccgttt
     1621 ccgtttccga tcgagcgaat cgatggttag gacccatggg ggacccatgg ggtgtcgtgt
     1681 ggtggtctgg tttgatccgc gatatttctc cgttcgtagt gtagatctga tcgaatccct
     1741 ggtgaaatcg ttgatcgtgc tattcgtgtg agggttctta ggtttggagt tgtggaggta
     1801 gttctgatcg gtttgtaggt gagattttcc ccatgatttt gcttggctcg tttgtcttgg
     1861 ttagattaga tctgcccgca ttttgttcga tatttctgat gcagatatga tgaataattt
     1921 cgtccttgta tcccgcgtcc gtatgtgtat taagtttgca ggtcctagtt aggtttttcc
     1981 tactgatttg tcttatccat tctgtttagc ttgcaaggtt tggtaatggt ccggcatgtt
     2041 tgtctctata gattagagta gaataagatt atctcaacaa gctgttggct tatcaatttt
     2101 ggatctgcat gtgtttcgca tctatatctt tgcaattaag atggtagatg gacatatgct
     2161 cctgttgagt tgatgttgta ccttttacct gaggtctgag gaacatgcat cctcctgcta
     2221 ctttgtgctt atacagatca tcaagattat gtagctaata ttcgatcagt ttctagtatc
     2281 tacatggtaa acttgcatgc acttgctact tatttttgat atacttggat gataacatat
     2341 gctgctggtt gattcctacc tacatgatga acattttaca ggccattagt gtctgtctgt
     2401 atgtgttgtt cctgtttgct tcagtctatt tctgtttcat tcctagttta ttggttctct
     2461 gctagatact taccctgctg ggcttagtta tcatcttatc tcgaatgcat tttcatgttt
     2521 atagatgaat atacactcag ataggtgtag atgtatgcta ctgtttctct acgttgctgt
     2581 aggttttacc tgtggcaact gcatactcct gttgcttcgc tagatatgta tgtgcttata
     2641 tagattaaga tatgtgtgat ggttctttag tatatctgat gatcatgtat gctcttttaa
     2701 cttcttgcta cacttggtaa catgctgtga tgctgtttgt tgattctgta gcactaccaa
     2761 tgatgacctt atctctcttt gtatatgatg tttctgtttg tttgaggctt gtgttactgc
     2821 tagttactta ccctgttgcc tggctaatct tctgcagatc ctgcaggaac aaagcaccag
     2881 tggtctagtg gtagaatagt accctgccac ggtacagacc cgggttcgat tcccggctgg
     2941 tgcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     3001 aagtggcacc gagtcggtgc aacaaagcac cagtggtcta gtggtagaat agtaccctgc
     3061 cacggtacag acccgggttc gattcccggc tggtgcaaag ccagataaca gtatgcgtat
     3121 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     3181 aaaggaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     3241 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     3301 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     3361 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     3421 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     3481 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     3541 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     3601 gcatgatgac caccgatatg gccagtgtgc cggtttccgt tatcggggaa gaagtggctg
     3661 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     3721 aagctcttca gtcgatcgac aagctcgagt ttctccataa taatgtgtga gtagttccca
     3781 gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct
     3841 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
     3901 aaatccagta ctaaaatcca gatcccccga attagagctc taccggtgac gcaggtgatg
     3961 aaaagcctta gggtcaggtg gcacttttcg gggaaatgtg cgatgaaccc ctatttgttt
     4021 atttttctaa atacattcaa atatgtatcc gctaatgaga caataaccct gataaatgct
     4081 tcaattatat tgaaaaagga agagtatgag tatccaacat ttccgtgtcg cccttattcc
     4141 cttttttgcg gccttctgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4201 agatgctgaa gatcagttgg gtgcccgagt gggttacatt gaactggatc tcaacagcgg
     4261 caaaatctta gagagttttc gccccgaaga acgcttccca atgatgagca ctttcaaagt
     4321 tctgctatgt ggcgcggtgt tatcccgtat tgatgctggg caagagcaac tcggtcgccg
     4381 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     4441 ggatggcatg acagtaagag aattatgtag tgctgccata accatgagtg ataacactgc
     4501 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     4561 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4621 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaactctcc gcaaactatt
     4681 aactggcgaa ctacttactc ttgcttcacg ccagcaactc attgactgga tggaggcgga
     4741 caaagttgca ggaccacttc tgcgctcggc acttccggct ggctggttta ttgctgataa
     4801 atctggagcc ggtgagcgtg gctctcgcgg tatcattgca gcactggggc cagatggtaa
     4861 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4921 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagacgaagg
     4981 gaaataaata gtagcccgcc tgatgtgcgg gctttttttt ggtggtggtt accagtgtcg
     5041 cttagctacc gtaggtcctc ttaggcggtc cgattagcga catagtcatc aat
//
