LOCUS       pBX_Spas_sgRNA_c        5562 bp ds-DNA     circular     01-OCT-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pBX Spas sgRNA BbsI Sequenced
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, May 21, 2016 from SnapGene 3.1.2
            http://www.snapgene.com
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     terminator      287..330
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /label=Escherichia coli rrnB
                     /ApEinfo_fwdcolor=#9d1b1c
                     /ApEinfo_revcolor=#9d1b1c
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     complement(473..489)
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label=SK primer
                     /ApEinfo_fwdcolor=#14c0bd
                     /ApEinfo_revcolor=#4ec02b
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        complement(522..540)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label=T7 promoter
                     /ApEinfo_fwdcolor=#346ee0
                     /ApEinfo_revcolor=#346ee0
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     complement(550..566)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label=M13 fwd
                     /ApEinfo_fwdcolor=#14c0bd
                     /ApEinfo_revcolor=#4ec02b
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             complement(1275..1937)
                     /codon_start=1
                     /product="replication protein for the broad-host-range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /note="pBBR1 Rep"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL
                     H
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYV
                     V
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGE
                     E
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
                     /label=replication protein for the broad-host-range
                     plasmid pBBR1 from Bordetella bronchiseptica
                     /ApEinfo_fwdcolor=#e9d024
                     /ApEinfo_revcolor=#e9d024
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      1938..2709
                     /note="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
                     /label=pBBR1 oriV
                     /ApEinfo_fwdcolor=#999999
                     /ApEinfo_revcolor=#999999
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             4189..4983
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     P
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     S
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     Q
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     A LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /label=aminoglycoside phosphotransferase from Tn5
                     /ApEinfo_fwdcolor=#e9d024
                     /ApEinfo_revcolor=#e9d024
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     terminator      5331..5377
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /label=rrnB T1 terminator
                     /ApEinfo_fwdcolor=#9d1b1c
                     /ApEinfo_revcolor=#9d1b1c
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    71..180
                     /label=pxyl
                     /ApEinfo_fwdcolor=#808000
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    201..281
                     /label=sgRNA
                     /ApEinfo_fwdcolor=cyan
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    181..200
                     /label=Seed region
                     /ApEinfo_fwdcolor=#ff8000
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 GAGCGCGGCC AGCAGGGCGG CTTCCTGGTC AGGGGTCAGG TCGGTCATGG GCAAGAGGTC
       61 CAGGTCGTGG TTTGTCGGCG GCTTCTAGCA TGGACCGCCC GCGCCCGTGA GGCCGAGGAT
      121 TTCGCGCTGG TCAGACAACC TACTTGCCGT CCCCACATGT TAGCGCTACC AAGTGCCGAC
      181 GTTAATTTAA GACTGGTTAA GTTTTTGTAC TCGAAAGAGC CTACAAAGAT AAGGCTTTAT
      241 GCCGAATTCA AGCACCCCAT GTTTTGACAT GAGGTGCTTT TTTTTGATAA AACGAAAGGC
      301 CCAGTCTTTC GACTGAGCCT TTCGTTTTAT AAACGGGTCT TGAGGGGTTT TTTAAGCTTC
      361 TGCAGCTGGA TCCATGGATA TCTAGATTTC CGCAAAATAA AAAAGGGGAC CTCTAGGGTC
      421 CCCAATTAAT TAGCTAAAGC CGCTAGCCCA TGGGAATTCC TGCAGCCCGG GGGATCCACT
      481 AGTTCTAGAG CGGCCGCCAC CGCGGTGGAG CTCCAATTCG CCCTATAGTG AGTCGTATTA
      541 CGCGCGCTCA CTGGCCGTCG TTTTACAACG TCGTGACTGG GAAAACCCTG GCGTTACCCA
      601 ACTTAATCGC CTTGCAGCAC ATCCCCCTTT CGCCAGCTGG CGTAATAGCG AAGAGGCCCG
      661 CACCGATCGC CCTTCCCAAC AGTTGCGCAG CCTGAATGGC GAATGGAAAT TGTAAGCGTT
      721 AATATTTTGT TAAAATTCGC GTTAAATTTT TGTTAAATCA GCTCATTTTT TAACCAATAG
      781 GCCGTACTGC GATGAGTGGC AGGGCGGGGC GTAATTTTTT TAAGGCAGTT ATTGGTGCCC
      841 TTAAACGCCT GGTTGCTACG CCTGAATAAG TGATAATAAG CGGATGAATG GCAGAAATTC
      901 GAAAGCAAAT TCGACCCGGT CGTCGGTTCA GGGCAGGGTC GTTAAATAGC CGCTTATGTC
      961 TATTGCTGGT TTACCGGTTT ATTGACTACC GGAAGCAGTG TGACCGTGTG CTTCTCAAAT
     1021 GCCTGAGGCC AGTTTGCTCA GGCTCTCCCC GTGGAGGTAA TAATTGACGA TATGATCATT
     1081 TATTCTGCCT CCCAGAGCCT GATAAAAACG GTGAATCCGT TAGCGAGGTG CCGCCGGCTT
     1141 CCATTCAGGT CGAGGTGGCC CGGCTCCATG CACCGCGACG CAACGCGGGG AGGCAGACAA
     1201 GGTATAGGGC GGCGAGGCGG CTACAGCCGA TAGTCTGGAA CAGCGCACTT ACGGGTTGCT
     1261 GCGCAACCCA AGTGCTACCG GCGCGGCAGC GTGACCCGTG TCGGCGGCTC CAACGGCTCG
     1321 CCATCGTCCA GAAAACACGG CTCATCGGGC ATCGGCAGGC GCTGCTGCCC GCGCCGTTCC
     1381 CATTCCTCCG TTTCGGTCAA GGCTGGCAGG TCTGGTTCCA TGCCCGGAAT GCCGGGCTGG
     1441 CTGGGCGGCT CCTCGCCGGG GCCGGTCGGT AGTTGCTGCT CGCCCGGATA CAGGGTCGGG
     1501 ATGCGGCGCA GGTCGCCATG CCCCAACAGC GATTCGTCCT GGTCGTCGTG ATCAACCACC
     1561 ACGGCGGCAC TGAACACCGA CAGGCGCAAC TGGTCGCGGG GCTGGCCCCA CGCCACGCGG
     1621 TCATTGACCA CGTAGGCCGA CACGGTGCCG GGGCCGTTGA GCTTCACGAC GGAGATCCAG
     1681 CGCTCGGCCA CCAAGTCCTT GACTGCGTAT TGGACCGTCC GCAAAGAACG TCCGATGAGC
     1741 TTGGAAAGTG TTTTCTGGCT GACCACCACG GCGTTCTGGT GGCCCATCTG CGCCACGAGG
     1801 TGATGCAGCA GCATTGCCGC CGTGGGTTTC CTCGCAATAA GCCCGGCCCA CGCCTCATGC
     1861 GCTTTGCGTT CCGTTTGCAC CCAGTGACCG GGCTTGTTCT TGGCTTGAAT GCCGATTTCT
     1921 CTGGACTGCG TGGCCATGCT TATCTCCATG CGGTAGGGGT GCCGCACGGT TGCGGCACCA
     1981 TGCGCAATCA GCTGCAACTT TTCGGCAGCG CGACAACAAT TATGCGTTGC GTAAAAGTGG
     2041 CAGTCAATTA CAGATTTTCT TTAACCTACG CAATGAGCTA TTGCGGGGGG TGCCGCAATG
     2101 AGCTGTTGCG TACCCCCCTT TTTTAAGTTG TTGATTTTTA AGTCTTTCGC ATTTCGCCCT
     2161 ATATCTAGTT CTTTGGTGCC CAAAGAAGGG CACCCCTGCG GGGTTCCCCC ACGCCTTCGG
     2221 CGCGGCTCCC CCTCCGGCAA AAAGTGGCCC CTCCGGGGCT TGTTGATCGA CTGCGCGGCC
     2281 TTCGGCCTTG CCCAAGGTGG CGCTGCCCCC TTGGAACCCC CGCACTCGCC GCCGTGAGGC
     2341 TCGGGGGGCA GGCGGGCGGG CTTCGCCCTT CGACTGCCCC CACTCGCATA GGCTTGGGTC
     2401 GTTCCAGGCG CGTCAAGGCC AAGCCGCTGC GCGGTCGCTG CGCGAGCCTT GACCCGCCTT
     2461 CCACTTGGTG TCCAACCGGC AAGCGAAGCG CGCAGGCCGC AGGCCGGAGG CTTTTCCCCA
     2521 GAGAAAATTA AAAAAATTGA TGGGGCAAGG CCGCAGGCCG CGCAGTTGGA GCCGGTGGGT
     2581 ATGTGGTCGA AGGCTGGGTA GCCGGTGGGC AATCCCTGTG GTCAAGCTCG TGGGCAGGCG
     2641 CAGCCTGTCC ATCAGCTTGT CCAGCAGGGT TGTCCACGGG CCGAGCGAAG CGAGCCAGCC
     2701 GGTGGCCGCT CGCGGCCATC GTCCACATAT CCACGGGCTG GCAAGGGAGC GCAGCGACCG
     2761 CGCAGGGCGA AGCCCGGAGA GCAAGCCCGT AGGGCGCCGC AGCCGCCGTA GGCGGTCACG
     2821 ACTTTGCGAA GCAAAGTCTA GTGAGTATAC TCAAGCATTG AGTGGCCCGC CGGAGGCACC
     2881 GCCTTGCGCT GCCCCCGTCG AGCCGGTTGG ACACCAAAAG GGAGGGGCAG GCATGGCGGC
     2941 ATACGCGATC ATGCGATGCA AGAAGCTGGC GAAAATGGGC AACGTGGCGG CCAGTCTCAA
     3001 GCACGCCTAC CGCGAGCGCG AGACGCCCAA CGCTGACGCC AGCAGGACGC CAGAGAACGA
     3061 GCACTGGGCG GCCAGCAGCA CCGATGAAGC GATGGGCCGA CTGCGCGAGT TGCTGCCAGA
     3121 GAAGCGGCGC AAGGACGCTG TGTTGGCGGT CGAGTACGTC ATGACGGCCA GCCCGGAATG
     3181 GTGGAAGTCG GCCAGCCAAG AACAGCAGGC GGCGTTCTTC GAGAAGGCGC ACAAGTGGCT
     3241 GGCGGACAAG TACGGGGCGG ATCGCATCGT GACGGCCAGC ATCCACCGTG ACGAAACCAG
     3301 CCCGCACATG ACCGCGTTCG TGGTGCCGCT GACGCAGGAC GGCAGGCTGT CGGCCAAGGA
     3361 GTTCATCGGC AACAAAGCGC AGATGACCCG CGACCAGACC ACGTTTGCGG CCGCTGTGGC
     3421 CGATCTAGGG CTGCAACGGG GCATCGAGGG CAGCAAGGCA CGTCACACGC GCATTCAGGC
     3481 GTTCTACGAG GCCCTGGAGC GGCCACCAGT GGGCCACGTC ACCATCAGCC CGCAAGCGGT
     3541 CGAGCCACGC GCCTATGCAC CGCAGGGATT GGCCGAAAAG CTGGGAATCT CAAAGCGCGT
     3601 TGAGACGCCG GAAGCCGTGG CCGACCGGCT GACAAAAGCG GTTCGGCAGG GGTATGAGCC
     3661 TGCCCTACAG GCCGCCGCAG GAGCGCGTGA GATGCGCAAG AAGGCCGATC AAGCCCAAGA
     3721 GACGGCCCGA GACCTTCGGG AGCGCCTGAA GCCCGTTCTG GACGCCCTGG GGCCGTTGAA
     3781 TCGGGATATG CAGGCCAAGG CCGCCGCGAT CATCAAGGCC GTGGGCGAAA AGCTGCTGAC
     3841 GGAACAGCGG GAAGTCCAGC GCCAGAAACA GGCCCAGCGC CAGCAGGAAC GCGGGCGCGC
     3901 ACATTTCCCC GAAAAGTGCC ACCTGGGATG AATGTCAGCT ACTGGGCTAT CTGGACAAGG
     3961 GAAAACGCAA GCGCAAAGAG AAAGCAGGTA GCTTGCAGTG GGCTTACATG GCGATAGCTA
     4021 GACTGGGCGG TTTTATGGAC AGCAAGCGAA CCGGAATTGC CAGCTGGGGC GCCCTCTGGT
     4081 AAGGTTGGGA AGCCCTGCAA AGTAAACTGG ATGGCTTTCT TGCCGCCAAG GATCTGATGG
     4141 CGCAGGGGAT CAAGATCTGA TCAAGAGACA GGATGAGGAT CGTTTCGCAT GATTGAACAA
     4201 GATGGATTGC ACGCAGGTTC TCCGGCCGCT TGGGTGGAGA GGCTATTCGG CTATGACTGG
     4261 GCACAACAGA CAATCGGCTG CTCTGATGCC GCCGTGTTCC GGCTGTCAGC GCAGGGGCGC
     4321 CCGGTTCTTT TTGTCAAGAC CGACCTGTCC GGTGCCCTGA ATGAACTGCA GGACGAGGCA
     4381 GCGCGGCTAT CGTGGCTGGC CACGACGGGC GTTCCTTGCG CAGCTGTGCT CGACGTTGTC
     4441 ACTGAAGCGG GAAGGGACTG GCTGCTATTG GGCGAAGTGC CGGGGCAGGA TCTCCTGTCA
     4501 TCTCACCTTG CTCCTGCCGA GAAAGTATCC ATCATGGCTG ATGCAATGCG GCGGCTGCAT
     4561 ACGCTTGATC CGGCTACCTG CCCATTCGAC CACCAAGCGA AACATCGCAT CGAGCGAGCA
     4621 CGTACTCGGA TGGAAGCCGG TCTTGTCGAT CAGGATGATC TGGACGAAGA GCATCAGGGG
     4681 CTCGCGCCAG CCGAACTGTT CGCCAGGCTC AAGGCGCGCA TGCCCGACGG CGAGGATCTC
     4741 GTCGTGACCC ATGGCGATGC CTGCTTGCCG AATATCATGG TGGAAAATGG CCGCTTTTCT
     4801 GGATTCATCG ACTGTGGCCG GCTGGGTGTG GCGGACCGCT ATCAGGACAT AGCGTTGGCT
     4861 ACCCGTGATA TTGCTGAAGA GCTTGGCGGC GAATGGGCTG ACCGCTTCCT CGTGCTTTAC
     4921 GGTATCGCCG CTCCCGATTC GCAGCGCATC GCCTTCTATC GCCTTCTTGA CGAGTTCTTC
     4981 TGAGCGGGAC TCTGGGGTTC GAAATGACCG ACCAAGCGAC GCCCAACCTG CCATCACGAG
     5041 ATTTCGATTC CACCGCCGCC TTCTATGAAA GGTTGGGCTT CGGAATCGTT TTCCGGGACG
     5101 CCGGCTGGAT GATCCTCCAG CGCGGGGATC TCATGCTGGA GTTCTTCGCC CACCCCCATG
     5161 GGCAAATATT ATACGCAAGG CGACAAGGTG CTGATGCCGC TGGCGATTCA GGTTCATCAT
     5221 GCCGTTTGTG ATGGCTTCCA TGTCGGCAGA ATGCTTAATG AATTACAACA GTTTTTATAT
     5281 AAGCTTTCAG TCGGCGCGAA TGCTCCACGC CGCGCCCAGC GGCGCGGATC CAAATAAAAC
     5341 GAAAGGCTCA GTCGAAAGAC TGGGCCTTTC GTTTTATCTG TTGTTTGTCG CTGCAGCCAG
     5401 CCGTGGTCGG GCAGCAGGTA GAAGGCGCCC TCGTCCTGAT CCTCGCCCGA AACCTCCAGC
     5461 CCCCGGTCGA TGGCTTCGAC GACATAGCCG GCCGCGCGGC AGGTGTCGGT GAGCGCGGCC
     5521 AGCAGGGCGG CTTCCTGGTC AGGGGTCAGG TCGGTCATGG GC 
//
