LOCUS       pBX_The3_sgRNA_g        5511 bp ds-DNA     circular     01-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBX The3 BbsI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 19, 2016 from SnapGene 3.1.0
            http://www.snapgene.com
COMMENT     
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     CDS             458..1252
                     /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"
                     /locus_tag="aminoglycoside phosphotransferase from Tn5"
                     /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      1600..1646
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /locus_tag="rrnB T1 terminator"
                     /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    1979..2064
                     /note="The3 sgRNA"
                     /locus_tag="The3 sgRNA"
                     /label=The3 sgRNA
                     /ApEinfo_fwdcolor=#7eff74
                     /ApEinfo_revcolor=#7eff74
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     terminator      2070..2113
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /locus_tag="Escherichia coli rrnB"
                     /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(2256..2272)
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /locus_tag="SK primer"
                     /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(2305..2323)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /locus_tag="T7 promoter"
                     /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(2333..2349)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /locus_tag="M13 fwd"
                     /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(3057..3719)
                     /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"
                     /locus_tag="replication protein for the broad-host-range
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /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      3720..4489
                     /note="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
                     /locus_tag="pBBR1 oriV"
                     /label=pBBR1 oriV
                     /ApEinfo_fwdcolor=#999999
                     /ApEinfo_revcolor=#999999
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1661..1959
                     /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    1960..1978
                     /label=guided sequence_gcrA_8
                     /ApEinfo_fwdcolor=cyan
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
      241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
      301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
      361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatata agctttcagt
     1561 cggcgcgaat gctccacgcc gcgcccagcg gcgcggatcc aaataaaacg aaaggctcag
     1621 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgc tgcagccagc cgtggtcggg
     1681 cagcaggtag aaggcgccct cgtcctgatc ctcgcccgaa acctccagcc cccggtcgat
     1741 ggcttcgacg acatagccgg ccgcgcggca ggtgtcggtg agcgcggcca gcagggcggc
     1801 ttcctggtca ggggtcaggt cggtcatggg caagaggtcc aggtcgtggt ttgtcggcgg
     1861 cttctagcat ggaccgcccg cgcccgtgag gccgaggatt tcgcgctggt cagacaacct
     1921 acttgccgtc cccacatgtt agcgctacca agtgccgacG GATCGATCAC CATATATAgt
     1981 tttagagctg tgaaaacagc gagttaaaat aaggcttagt ccgtactcaa cttgaaaagg
     2041 tggcaccgat tcggtgtttt ttttttttga taaaacgaaa ggcccagtct ttcgactgag
     2101 cctttcgttt tataaacggg tcttgagggg ttttttaagc ttctgcagct ggatccatgg
     2161 atatctagat ttccgcaaaa taaaaaaggg gacctctagg gtccccaatt aattagctaa
     2221 agccgctagc ccatgggaat tcctgcagcc cgggggatcc actagttcta gagcggccgc
     2281 caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     2341 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2401 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2461 aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt
     2521 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgtac tgcgatgagt
     2581 ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggtgcta
     2641 cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg
     2701 gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt
     2761 ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct
     2821 caggctctcc ccgtggaggt aataattgac gatatgatca tttattctgc ctcccagagc
     2881 ctgataaaaa cggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     2941 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgaggc
     3001 ggctacagcc gatagtctgg aacagcgcac ttacgggttg ctgcgcaacc caagtgctac
     3061 cggcgcggca gcgtgacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac
     3121 ggctcatcgg gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc
     3181 aaggctggca ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg
     3241 gggccggtcg gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca
     3301 tgccccaaca gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc
     3361 gacaggcgca actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc
     3421 gacacggtgc cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc
     3481 ttgactgcgt attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgttttctgg
     3541 ctgaccacca cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc
     3601 gccgtgggtt tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc
     3661 acccagtgac cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg
     3721 cttatctcca tgcggtaggg tgccgcacgg ttgcggcacc atgcgcaatc agctgcaact
     3781 tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg gcagtcaatt acagattttc
     3841 tttaacctac gcaatgagct attgcggggg gtgccgcaat gagctgttgc gtacccccct
     3901 tttttaagtt gttgattttt aagtctttcg catttcgccc tatatctagt tctttggtgc
     3961 ccaaagaagg gcacccctgc ggggttcccc cacgccttcg gcgcggctcc ccctccggca
     4021 aaaagtggcc cctccggggc ttgttgatcg actgcgcggc cttcggcctt gcccaaggtg
     4081 gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg ctcggggggc aggcgggcgg
     4141 gcttcgcctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc
     4201 aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg tccaaccggc
     4261 aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta aaaaaattga
     4321 tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga aggctgggta
     4381 gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt
     4441 ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct cgcggccatc
     4501 gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga agcccggaga
     4561 gcaagcccgt agggcgccgc agccgccgta ggcggtcacg actttgcgaa gcaaagtcta
     4621 gtgagtatac tcaagcattg agtggcccgc cggaggcacc gccttgcgct gcccccgtcg
     4681 agccggttgg acaccaaaag ggaggggcag gcatggcggc atacgcgatc atgcgatgca
     4741 agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac cgcgagcgcg
     4801 agacgcccaa cgctgacgcc agcaggacgc cagagaacga gcactgggcg gccagcagca
     4861 ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc aaggacgctg
     4921 tgttggcggt cgagtacgtc atgacggcca gcccggaatg gtggaagtcg gccagccaag
     4981 aacagcaggc ggcgttcttc gagaaggcgc acaagtggct ggcggacaag tacggggcgg
     5041 atcgcatcgt gacggccagc atccaccgtg acgaaaccag cccgcacatg accgcgttcg
     5101 tggtgccgct gacgcaggac ggcaggctgt cggccaagga gttcatcggc aacaaagcgc
     5161 agatgacccg cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg ctgcaacggg
     5221 gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc gttctacgag gccctggagc
     5281 ggccaccagt gggccacgtc accatcagcc cgcaagcggt cgagccacgc gcctatgcac
     5341 cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg gaagccgtgg
     5401 ccgaccggct gacaaaagcg gttcggcagg ggtatgagcc tgccctacag gccgccgcag
     5461 gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga gacggcccga g
//
