LOCUS       Exported                2349 bp ds-DNA     circular SYN 01-FEB-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    psgRNAc BsaI
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 2349)
  AUTHORS   Bikard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 24, 2015 from SnapGene 2.7.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2349
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        complement(220..322)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /note="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the colE1 origin."
     CDS             1467..1482
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /note="Cas9"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="LGGD"
     promoter        1510..1538
                     /note="Promoter (BBa_J23119)"
                     /note="from BioBricks"
     -35_signal      1510..1515
                     /note="-35"
     -10_signal      1533..1538
                     /note="-10"
     misc_feature    1545..1564
                     /note="Control spacer with 2 BsaI"
     misc_RNA        1565..1640
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the CRISPR/Cas9 system"
     misc_feature    1642..1647
                     /note="part of the tracrRNA"
     CDS             1648..1652
                     /codon_start=1
                     /transl_table=11
                     /gene="lacI"
                     /gene_synonym="ECK0342; JW0336"
                     /locus_tag="b0345"
                     /product="lactose-inducible lac operon transcriptional 
                     repressor"
                     /function="regulator; Degradation of small molecules: 
                     Carbon compounds"
                     /note="lacI"
                     /note="transcriptional repressor of the lac operon"
                     /db_xref="GI:49176012"
                     /db_xref="ASAP:ABE-0001189"
                     /db_xref="UniProtKB/Swiss-Prot:P03023"
                     /db_xref="EcoGene:EG10525"
                     /db_xref="GeneID:945007"
                     /experiment="N-terminus verified by Edman degradation: PMID
                     1107032,4554534,4557395,4571224"
                     /protein_id="NP_414879.3"
                     /translation="V"
     gene            1648..1652
                     /gene="lacI"
                     /gene_synonym="ECK0342; JW0336"
                     /locus_tag="b0345"
                     /note="lacI"
                     /db_xref="EcoGene:EG10525"
                     /db_xref="GeneID:945007"
     CDS             complement(1909..219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctggcta ggaggtgact gaagtatatt ttaggaattc
     1501 taaagatctt tgacagctag ctcagtccta ggtataatac tagttgagac cagtctaggt
     1561 ctcggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     1621 aagtggcacc gagtcggtgc tttttttggt agtgcagcgc gatcgtaatc aggggggaga
     1681 gcctgagcaa actggcctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     1741 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     1801 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     1861 aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     1921 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1981 acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     2041 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     2101 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     2161 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     2221 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     2281 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     2341 tgccatacg
//
