LOCUS       Exported                6135 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMDC21, complete sequence.
ACCESSION   KY406738
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6135)
  AUTHORS   Shao Y.
  TITLE     Establishment of markerless gene deletion system in Chromohalobacter
            salexigens
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6135)
  AUTHORS   Shao Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-DEC-2016) College of Life Science, Qingdao 
            Agricultural University, The Great Wall Road, No. 700, Qingdao, 
            Shandong 266109, China
REFERENCE   3  (bases 1 to 6135)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6135
                     /organism="Cloning vector pMDC21"
                     /mol_type="other DNA"
                     /db_xref="taxon:2009236"
     promoter        565..667
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             668..1327
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="ARX95167.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            668..1327
                     /gene="cat"
                     /label=cat
     promoter        1602..2047
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2048..3469
                     /codon_start=1
                     /gene="sacB"
                     /product="levan sucrase"
                     /label=sacB
                     /protein_id="ARX95168.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            2048..3469
                     /gene="sacB"
                     /label=sacB
     oriT            3823..3932
                     /note="incP origin of transfer"
     promoter        complement(4167..4257)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        complement(4451..4541)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4901..5495
                     /label=pMB1 replication origin
                     /note="pMB1 replication origin"
     rep_origin      4901..5489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5777..5798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5813..5843
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5851..5867
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5875..5891
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5880..5895
                     /note="pK18M13R; special M13R binding site"
     CDS             5892..6122
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="LacZ alpha fragment"
                     /label=lacZ alpha
                     /protein_id="ARX95169.1"
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            5892..6122
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     misc_feature    5900..5956
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5960..5977)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(5960..5976)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"