LOCUS       sequence-208548-        2786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of Sa sgRNA from the T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2786)
  AUTHORS   Liu Z, Lu Z, Yang G, Huang S, Li G, Feng S, Liu Y, Li J, Yu W, Zhang
            Y, Chen J, Sun Q, Huang X
  TITLE     Efficient generation of mouse models of human diseases via ABE- and 
            BE-mediated base editing.
  JOURNAL   Nat Commun. 2018 Jun 14;9(1):2338. doi: 10.1038/s41467-018-04768-7.
  PUBMED    29904106
REFERENCE   2  (bases 1 to 2786)
  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 
            Commun."; date: "2018-06-14"; volume: "9(1):2338. doi"; pages: " 
            10.1038/s41467-018-04768-7"
COMMENT     Sequence Label: pUC57-Sa sgRNA expression vector
FEATURES             Location/Qualifiers
     source          1..2786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(168..184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(168..184)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(181..203)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    192..208
                     /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)."
     promoter        complement(join(216..222,223..240,241..246))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    261..282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(400..417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(571..1159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(651..670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1330..2145)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1330..2145)
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2053..2072
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2146..2250)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2318..2336
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2374..2396)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2496..2515
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2709..2731
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2723..2740
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2724..2740
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggt gagaccgaga gagggtctca gttttagtac tctgtaatga
       61 aaattacaga atctactaaa acaaggcaaa atgccgtgtt tatctcgtca acttgttggc
      121 gagatttttt tctgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt
      181 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      241 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      301 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      361 gggagaggcg gtttgcgtat tgggcgcggc cgccgcttcc tcgctcactg actcgctgcg
      421 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      481 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      541 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1321 gtctgacagt tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     1381 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     1441 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     1501 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     1561 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     1621 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     1681 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     1741 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     1801 atcaggatat tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa
     1861 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     1921 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     1981 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     2041 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     2101 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca
     2161 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2221 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2281 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2341 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2401 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2461 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2521 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2581 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     2641 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     2701 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacctc
     2761 gcgaatgcat ctagatatcg gatccc
//
