LOCUS       sequence-211551-        4297 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Blasticidin resistant cassette for knock-in by VIKING method.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4297)
  AUTHORS   Sawatsubashi S, Joko Y, Fukumoto S, Matsumoto T, Sugano SS
  TITLE     Development of versatile non-homologous end joining-based knock-in 
            module for genome editing.
  JOURNAL   Sci Rep. 2018 Jan 12;8(1):593. doi: 10.1038/s41598-017-18911-9.
  PUBMED    29330493
REFERENCE   2  (bases 1 to 4297)
  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: "Sci Rep."; 
            date: "2018-01-12"; volume: "8(1):593. doi"; pages: " 
            10.1038/s41598-017-18911-9"
COMMENT     Sequence Label: pVKG1-BSD
FEATURES             Location/Qualifiers
     source          1..4297
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        1..317
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      168..303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          916..981
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1111..1131
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1556..1677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1593..1612)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1647..1666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1704..1803)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1820..1839)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1821..1839)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1844..1860)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1844..1860)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1962..1981)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1973..2782
                     /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     complement(2040..2059)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2929..3517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3418..3437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3671..3688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3847..3874)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3966..4052)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4101..4123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4115..4132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4116..4132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4148..4247
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtccca aagtccccag gctccccagc aggcagaagt atgcaaagca
      121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
      181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
      241 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      301 gcctaggctt ttgcaaaaag ctttctcatt gtcaatatgc ctttgtctca agaagaatcc
      361 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
      421 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
      481 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
      541 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
      601 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
      661 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
      721 gagggctaag caccagccat tgatcttgta cattcaagct actttctgtt actagatgaa
      781 ccgagcatga gttgatggtg actgcaggag tggggaggca cgatggccgc tttggtccgg
      841 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
      901 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt
      961 ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg
     1021 aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag
     1081 gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc
     1141 aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact
     1201 cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt
     1261 atggaaaaat attctgtaac ctttataagt aggcataaca gttataatca taacatactg
     1321 ttttttctta ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg
     1381 tgtaccttta gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc
     1441 ttgactagag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1501 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1561 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1621 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     1681 tgtctggaag ggtgggcgcg ccgacccagc tttcttgtac aaagttggca ttataagaaa
     1741 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1801 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1861 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1921 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     1981 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2041 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2101 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2161 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2221 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2281 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2341 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2401 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2461 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2521 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2581 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2641 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2701 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2761 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     2821 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     2881 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2941 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3001 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3061 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3121 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3181 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3241 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3301 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3361 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3421 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3481 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3541 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3601 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3661 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3721 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3781 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     3841 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3901 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3961 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     4021 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     4081 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     4141 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     4201 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctccg cggccgcccc
     4261 cttcaccgga tccggctgtg gaatgtgtgt cagttag
//
