LOCUS       sequence-319702-        2244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Version of SgRNA scaffold with the sequence 2.1 of Ms2 aptamer in 
            the 3' end..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2244)
  AUTHORS   Selma S, Bernabe-Orts JM, Vazquez-Vilar M, Diego-Martin B, Ajenjo M,
            Garcia-Carpintero V, Granell A, Orzaez D
  TITLE     Strong gene activation in plants with genome-wide specificity using 
            a new orthogonal CRISPR/Cas9-based programmable transcriptional 
            activator.
  JOURNAL   Plant Biotechnol J. 2019 Sep;17(9):1703-1705. doi: 
            10.1111/pbi.13138. Epub 2019 May 23.
  PUBMED    31034138
REFERENCE   2  (bases 1 to 2244)
  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: "10.1111/pbi.13138";
            journalName: "Plant Biotechnol J"; date: "2019-09"; volume: "17"; 
            issue: "9"; pages: "1703-1705"
COMMENT     Sequence Label: pUPD2 sgRNAscaffold 2.1 scRNA (GB1437)
FEATURES             Location/Qualifiers
     source          1..2244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(737..831)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(852..1511)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1426..1445
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1673..1691
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1695..1738)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1741..1762
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_RNA        1778..1853
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      1958..2015
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2039..2056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2210..2244,1..554))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
       61 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      601 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      661 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      721 acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
      781 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
      841 tcgatatcaa attacgcccc gccctgccac tcgtcgcagt actgttgtaa ttcattaagc
      901 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
      961 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     1021 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgatacg
     1081 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     1141 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagt
     1201 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     1261 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     1321 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     1381 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     1441 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     1501 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     1561 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaactcg
     1621 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1681 tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg
     1741 gaattcgcgg ccgcttctag agcgatgagt ggtctcggtt ttagagctag aaatagcaag
     1801 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcgggagca
     1861 catgaggaat cacccatgtg cgactcccac agtcactggg gagtcttccc ttttttcgct
     1921 tgagaccact catcgcgact agtagcggcc gctgcagtcc ggcaaaaaag ggcaaggtgt
     1981 caccaccctg ccctttttct ttaaaaccga aaagattact tcgcgttatg caggcttcct
     2041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccacaggc
     2221 tccgcccccc tgacgagcat caca
//
