LOCUS       sequence-209776-        2876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial sgRNA expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2876)
  AUTHORS   Banno S, Nishida K, Arazoe T, Mitsunobu H, Kondo A
  TITLE     Deaminase-mediated multiplex genome editing in Escherichia coli.
  JOURNAL   Nat Microbiol. 2018 Feb 5. pii: 10.1038/s41564-017-0102-6. doi: 
            10.1038/s41564-017-0102-6.
  PUBMED    29403014
REFERENCE   2  (bases 1 to 2876)
  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: "Nat 
            Microbiol. 2018 Feb 5. pii: 10.1038/s41564-017-0102-6. doi: 
            10.1038/s41564-017-0102-6."
COMMENT     Sequence Label: pTAKN2_J23119-sgRNA
FEATURES             Location/Qualifiers
     source          1..2876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     405..424
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        405..423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        499..533
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        554..629
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        709..743
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        764..839
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(932..949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1103..1691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1183..1202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1849..2664)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTSAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2572..2591
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2656..2675
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2849..2867
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaatattac tacaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgagct agtgtaatac gactcactat
      421 agggcgcggc cgcagaattc gataacaatt gagcaagaat cttcatcgat gcatcagaaa
      481 attattttaa atttcctctt gacagctagc tcagtcctag gtataatgct agcagagacc
      541 cgggatggtc tcagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      601 aacttgaaaa agtggcaccg agtcggtgct ttttttccga aaaaaaaacc ccgcccctga
      661 cagggcgggg ttttttttaa ttaacagaaa attattttaa atttcctctt gacagctagc
      721 tcagtcctag gtataatgct agcagaagag ccggcgctct tcagttttag agctagaaat
      781 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      841 ttttttccga aaaaaaaacc ccgcccctga cagggcgggg tttttttttc tagaccgcaa
      901 aaatcagcgc gcaaatacgc atacaagctt cctcgctcac tgactcgctg cgctcggtcg
      961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgttttaatt agaaaaactc
     1861 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     1921 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     1981 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     2041 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     2101 gaatggcaat agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     2161 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     2221 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     2281 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     2341 ctggaatgct gttttccctg ggatcgcaga ggtgagtaac catgcatcat caggagtacg
     2401 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     2461 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     2521 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     2581 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc ttgagcaaga
     2641 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacat
     2701 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2761 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2821 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
