LOCUS       sequence-252876-        2543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  classic sgRNA backbone for tRNA-gRNA array, Position 1, combine with
            TaU3 promoter module (pFH31).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2543)
  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 2543)
  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: pAK007
FEATURES             Location/Qualifiers
     source          1..2543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    217..238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(253..258,259..276,277..283)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    291..307
                     /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     296..318
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     315..331
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     315..331
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             327..650
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    344..400
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(401..418)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(401..417)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(410..432)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(626..645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(855..1443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(935..954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1621..2430)
                     /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     2344..2363
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2422..2441
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gacgtcattg
       61 tcgatggtct caaagcaaca aagcaccagt ggtctagtgg tagaatagta ccctgccacg
      121 gtacagaccc gggttcgatt cccggctggt gcattgtctt cgcagctggc acgacaggtt
      181 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      241 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      301 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
      361 ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
      421 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      481 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      541 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      601 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      661 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      721 cttacagaca agctgtgacg aagacaagtt ttagagctag aaatagcaag ttaaaattga
      781 gaccatcatt gtcacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      841 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      901 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      961 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1021 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1081 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1141 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1201 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1261 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1321 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1381 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1441 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1501 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1561 aaatgaagtt ttaaatcaag cccaatctga ataatgttac aaccaattaa ccaattctga
     1621 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     1681 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     1741 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     1801 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     1861 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     1921 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     1981 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     2041 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     2101 ttcttctaat acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc
     2161 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     2221 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     2281 caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac
     2341 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     2401 cctcgacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     2461 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     2521 ttgagacacg ggccagagct gca
//
