LOCUS       sequence-319685-        2248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A version of scaffold sgRNA with the sequence of MS2 aptamer inside 
            the tetraloop of the scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2248)
  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 2248)
  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 sgRNA:scaffold tetraloop MS2 aptamer SAM...
FEATURES             Location/Qualifiers
     source          1..2248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        18..36
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        88..106
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      185..242
                     /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(266..283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(437..1025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(517..536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1208..1302)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1323..1982)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1897..1916
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2144..2162
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2166..2209)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2212..2233
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gttttagagc taggccaaca tgaggatcac ccatgtctgc acggcctagc aagttaaaat
       61 aaggctagtc cgttatcaac ttggccaaca tgaggatcac ccatgtctgc acggccaagt
      121 ggcaccgagt cggtgctttt tttcgcttga gaccactcat cgcgactagt agcggccgct
      181 gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
      241 gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      301 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      361 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      421 ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac aaaaatcgac
      481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      841 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      901 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      961 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1081 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1141 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagctcga
     1201 ggcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     1261 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca
     1321 aattacgccc cgccctgcca ctcgtcgcag tactgttgta attcattaag cattctgccg
     1381 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     1441 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     1501 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgatac gaaaaacata
     1561 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     1621 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag tgatgaaaac
     1681 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     1741 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
     1801 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     1861 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     1921 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     1981 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     2041 cttatttcat tatggtgaaa gttggaacct cttacgtgcc cgatcaactc gagtgccacc
     2101 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2161 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg
     2221 gccgcttcta gagcgatgag tggtctcg
//
