LOCUS       Exported                4323 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Blasticidin resistant cassette for knock-in by VIKING method.
ACCESSION   .
VERSION     .
KEYWORDS    pVKG1-BSD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4323)
  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 4323)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4323
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             680..700
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(722..742)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        727..1056
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      907..1042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     969..988
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          1656..1721
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1851..1871
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2296..2430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2333..2352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2387..2406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2448..2547)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2564..2583)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2565..2583)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2588..2604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2588..2604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2706..2725)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2717..3526
                     /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(2784..2803)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3673..4261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4162..4181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgcct tgtttaactt taagaaggag cccttcaccg gatccggctg
      721 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      781 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
      841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      901 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      961 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     1021 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tttctcattg tcaatatgcc
     1081 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     1141 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     1201 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     1261 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     1321 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     1381 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     1441 gccctctggt tatgtgtggg agggctaagc accagccatt gatcttgtac attcaagcta
     1501 ctttctgtta ctagatgaac cgagcatgag ttgatggtga ctgcaggagt ggggaggcac
     1561 gatggccgct ttggtcccgg atctttgtga aggaacctta cttctgtggt gtgacataat
     1621 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     1681 aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg
     1741 atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa
     1801 tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga
     1861 agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg
     1921 ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg
     1981 cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt aggcataaca
     2041 gttataatca taacatactg ttttttctta ctccacacag gcatagagtg tctgctatta
     2101 ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg gttaataagg
     2161 aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag
     2221 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2281 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2401 ctcatcaatg tatcttatca tgtctggatc caagggtggg cgcgccgacc cagctttctt
     2461 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2521 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2581 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2641 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2701 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2761 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2821 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2881 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2941 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3001 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3061 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3121 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3181 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3241 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3301 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3361 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3421 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3481 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3541 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3601 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3661 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3721 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3781 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3841 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3901 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3961 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4021 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     4081 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4141 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4201 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4261 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4321 gtt
//
