LOCUS       sequence-252903-        2552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  improved sgRNA backbone for tRNA-gRNA array, Position 1, combine 
            with AtU6-26 promoter module (pFH34).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2552)
  AUTHORS   Hahn F, Korolev A, Sanjurjo Loures L, Nekrasov V
  TITLE     A modular cloning toolkit for genome editing in plants.
  JOURNAL   BMC Plant Biol. 2020 Apr 23;20(1):179. doi: 
            10.1186/s12870-020-02388-2.
  PUBMED    32326901
REFERENCE   2  (bases 1 to 2552)
  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: "BMC Plant 
            Biol."; date: "2020-04-23"; volume: "20(1):179. doi"; pages: " 
            10.1186/s12870-020-02388-2"
COMMENT     Sequence Label: pFH49
FEATURES             Location/Qualifiers
     source          1..2552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    216..237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(252..257,258..275,276..282)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    290..306
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     295..317
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     314..330
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     314..330
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             326..649
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    343..399
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(400..417)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(400..416)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(409..431)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(625..644)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(864..1452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(944..963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1630..2439)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2353..2372
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2431..2450
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tctggtctca gattgaacaa agcaccagtg gtctagtggt agaatagtac cctgccacgg
      121 tacagacccg ggttcgattc ccggctggtg cattgtcttc gcagctggca cgacaggttt
      181 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      241 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      301 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
      361 gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg
      421 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      481 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      541 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      601 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      661 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      721 ttacagacaa gctgtgacga agacaagttt cagagctatg ctggaaacag catagcaagt
      781 tgaaattgag accatcattg tcacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1561 ttaaattaaa aatgaagttt taaatcaagc ccaatctgaa taatgttaca accaattaac
     1621 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     1681 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     1741 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     1801 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     1861 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     1921 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     1981 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2041 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2101 atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa
     2161 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     2221 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     2281 tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga
     2341 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     2401 taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     2461 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     2521 tcagagattt tgagacacgg gccagagctg ca
//
