LOCUS       Exported                2541 bp ds-DNA     circular SYN 28-AUG-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2541)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 3, 2018 from SnapGene 4.1.4
            http://www.snapgene.com
COMMENT     LOCUS pgRNA-bacteria 2584 bp DNA SYN DEFINITION pgRNA-bacteria 
            ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial 
            sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..2541
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          51..70
                     /organism="Escherichia coli str. K-12 substr. MG1655"
                     /strain="K-12"
                     /sub_strain="MG1655"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:511145"
     promoter        16..44
                     /label=J23119 with SpeI immediately following
                     /note="J23119 with SpeI immediately following -10 hexemer"
     -35_signal      16..21
                     /label=-35
                     /note="-35"
     primer_bind     complement(20..72)
                     /label=gCRM052
     -10_signal      39..44
                     /label=-10
                     /note="-10"
     misc_feature    45..50
                     /label=SpeI site
                     /note="SpeI site"
     primer_bind     49..103
                     /label=gCRM051
     misc_feature    51..70
                     /label=gRNA
     CDS             complement(51..70)
                     /codon_start=1
                     /transl_table=11
                     /gene="pta"
                     /gene_synonym="ECK2291; JW2294"
                     /locus_tag="b2297"
                     /product="phosphate acetyltransferase"
                     /function="enzyme; Degradation of small molecules: Carbon 
                     compounds"
                     /EC_number="2.3.1.8"
                     /label=pta
                     /note="phosphotransacetylase"
                     /db_xref="GI:16130232"
                     /db_xref="ASAP:ABE-0007582"
                     /db_xref="UniProtKB/Swiss-Prot:P0A9M8"
                     /db_xref="EcoGene:EG20173"
                     /db_xref="GeneID:946778"
                     /protein_id="NP_416800.1"
                     /translation="TRKHQF"
     gene            complement(51..70)
                     /gene="pta"
                     /gene_synonym="ECK2291; JW2294"
                     /locus_tag="b2297"
                     /label=pta
                     /db_xref="EcoGene:EG20173"
                     /db_xref="GeneID:946778"
     misc_RNA        71..146
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the CRISPR/Cas9 system"
     misc_feature    71..112
                     /label=cas9 handle
                     /note="cas9 handle"
     misc_feature    113..153
                     /label=S. pyogenes terminator
                     /note="S. pyogenes terminator"
     misc_feature    154..254
                     /label=Spacer Included in Qi et. al. 2013
                     /note="Spacer Included in Qi et. al. 2013 ""Repurposing 
                     CRISPR..."""
     terminator      255..520
                     /label=rrnB_terminator
     misc_feature    complement(285..302)
                     /label=pBAD_rev_primer
     misc_feature    complement(285..302)
                     /label=pTrcHis_rev_primer
     terminator      455..500
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      458..501
                     /label=rrnB_T1_terminator
     rep_origin      complement(725..1313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1484..2299)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    complement(2358)
                     /label=TSP
                     /note="TSP"
     misc_feature    complement(2359..2394)
                     /label=cmR promoter
                     /note="cmR promoter"
     terminator      2496..2539
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt caaactggtg
       61 cttacgtgtc gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      121 ttgaaaaagt ggcaccgagt cggtgctttt tttgaagctt gggcccgaac aaaaactcat
      181 ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt gagtttaaac
      241 ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
      301 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
      361 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
      421 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
      481 agactgggcc tttcgtttta tctgttgttt gtcggtgaac tggatcctta ctcgagtcta
      541 gactgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1141 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     1201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagtcagaaa aactcatcga
     1501 gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1561 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1621 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1681 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1741 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1801 caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     1861 atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     1921 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     1981 atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2041 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg accatctcat
     2101 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2161 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2221 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     2281 cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2341 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtaggggt
     2401 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     2461 attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc
     2521 tttcgctaag gatgatttct g
//
