LOCUS       Exported                5247 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAC-crRNA-Cm, complete sequence.
ACCESSION   MF287368
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5247)
  AUTHORS   Yan MY, Yan HQ, Ren GX, Zhao JP, Guo XP, Sun YC.
  TITLE     CRISPR-Cas12a-assisted recombineering in bacteria
  JOURNAL   Appl. Environ. Microbiol. (2017) In press
  PUBMED    28646112
REFERENCE   2  (bases 1 to 5247)
  AUTHORS   Yan M-Y., Yan H-Q., Ren G-X., Zhao J-P., Guo X-P., Sun Y-C.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2017) Moh Key Laboratory of Systems Biology of 
            Pathogens, Chinese Academy of Medical Sciences and Peking Union 
            Medical College Institute of Pathogen Biology, 9, Dongdan Santiao, 
            Dongcheng District, Beijing 100730, China
REFERENCE   3  (bases 1 to 5247)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5247
                     /organism="Expression vector pAC-crRNA-Cm"
                     /mol_type="other DNA"
                     /db_xref="taxon:2018568"
     CDS             complement(1..660)
                     /codon_start=1
                     /gene="CAT"
                     /product="chloramphenicol acetyltransferase"
                     /label=CAT
                     /protein_id="ASK86108.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(1..660)
                     /gene="CAT"
                     /label=CAT
     regulatory      complement(661..763)
                     /regulatory_class="promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        complement(661..763)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1289..1834)
                     /direction=LEFT
                     /label=p15Aori
                     /note="p15Aori"
     promoter        1946..1974
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_feature    2003..2038
                     /label=Direct Repeat
                     /note="Direct Repeat"
     CDS             2058..2777
                     /codon_start=1
                     /product="AcGFP1"
                     /label=AcGFP1
                     /note="Aequorea coerulescens GFP"
                     /protein_id="ASK86109.1"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    2798..2833
                     /label=Direct Repeat
                     /note="Direct Repeat"
     regulatory      2937..3382
                     /regulatory_class="promoter"
                     /note="sacB promoter"
     promoter        2937..3382
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3383..4804
                     /codon_start=1
                     /product="sacB"
                     /label=sacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /protein_id="ASK86110.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"