LOCUS       Exported                2849 bp ds-DNA     circular SYN 11-JAN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    P167_pTGC-U6.3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 11, 2019 from SnapGene 3.3.2
            http://www.snapgene.com
COMMENT     pDONR221
FEATURES             Location/Qualifiers
     source          1..2849
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      complement(268..295)
                     /note="rrnB T2 transcription termination"
                     /note="rrnB T2 transcription termination sequence"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        576..651
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        653..1073
                     /note="dU6-3 promoter"
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     promoter        complement(1091..1109)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1114..1130)
                     /note="M13 reverse primer"
     CDS             1243..2052
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2199..2787
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 tcgggccgct cttctgtttt agagctagaa atagcaagtt
      601 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgc
      661 tcacctgtga ttgctcctac tcaaatacaa aaacatcaaa ttttctgtca ataaagcata
      721 tttatttata tttattttac aggaaagaat tccttttaaa gtgtatttta acctataatg
      781 aaaaacgatt aaaaaaaata cataaaataa ttcgaaaatt tttgaatagc ccaggttgat
      841 aaaaattcat ttcatacgtt ttataactta tgcccctaag tattttttga ccatagtgtt
      901 tcaattctac attaatttta cagagtagaa tgaaacgcca cctactcagc caagaggcga
      961 aaaggttagc tcgccaagca gagagggcgc cagtgctcac tactttttat aattctcaac
     1021 ttctttttcc agactcagtt cgtatatata gacctatttt caatttaacg tcgggaagag
     1081 cctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1141 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaataatatc atcatgaaca
     1201 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1261 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1321 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     1381 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1441 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1501 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     1561 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1621 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1681 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1741 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     1801 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1861 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     1921 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     1981 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2041 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2101 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2821 gccttttgct ggccttttgc tcacatgtt
//
