LOCUS       40924_30430        6135 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMDC21, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6135)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-DEC-2016) College of Life Science, Qingdao Agricultural 
            University, The Great Wall Road, No. 700, Qingdao, Shandong 266109, 
            China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        565..667
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             668..1324
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        1602..2047
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2048..3466
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     oriT            complement(3823..3932)
                     /direction=LEFT
                     /label=oriT
                     /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..5489
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5777..5798
                     /label=CAP binding site
                     /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
                     /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"
     misc_feature    5900..5956
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5960..5976)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
       61 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
      121 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
      181 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
      241 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
      301 aggggatcat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt agctgaacag
      361 gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca tcatacacta
      421 aatcagtaag ttggcagcat cacccgacgc actttgcgcc gaataaatac ctgtgacgga
      481 agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga agccctgggc
      541 caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa
      601 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
      661 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
      721 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
      781 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
      841 ctttattcac attcttgccc gcctgatgaa tgctcatccg gagttccgta tggcaatgaa
      901 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
      961 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1021 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1081 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1141 aaacgtggcc aatatggaca acttcttcgc ccccgttttc actatgggca aatattatac
     1201 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
     1261 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1321 ggcgtaacga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     1561 gactctgggg ttcgctagag gatcgatcct ttttaaccca tcacatatac ctgccgttca
     1621 ctattattta gtgaaatgag atattatgat attttctgaa ttgtgattaa aaaggcaact
     1681 ttatgcccat gcaacagaaa ctataaaaaa tacagagaat gaaaagaaac agatagattt
     1741 tttagttctt taggcccgta gtctgcaaat ccttttatga ttttctatca aacaaaagag
     1801 gaaaatagac cagttgcaat ccaaacgaga gtctaataga atgaggtcga aaagtaaatc
     1861 gcgcgggttt gttactgata aagcaggcaa gacctaaaat gtgtaaaggg caaagtgtat
     1921 actttggcgt caccccttac atattttagg tcttttttta ttgtgcgtaa ctaacttgcc
     1981 atcttcaaac aggagggctg gaagaagcag accgctaaca cagtacataa aaaaggagac
     2041 atgaacgatg aacatcaaaa agtttgcaaa acaagcaaca gtattaacct ttactaccgc
     2101 actgctggca ggaggcgcaa ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa
     2161 ggaaacatac ggcatttccc atattacacg ccatgatatg ctgcaaatcc ctgaacagca
     2221 aaaaaatgaa aaatatcaag tttctgaatt tgattcgtcc acaattaaaa atatctcttc
     2281 tgcaaaaggc ctggacgttt gggacagctg gccattacaa aacgctgacg gcactgtcgc
     2341 aaactatcac ggctaccaca tcgtctttgc attagccgga gatcctaaaa atgcggatga
     2401 cacatcgatt tacatgttct atcaaaaagt cggcgaaact tctattgaca gctggaaaaa
     2461 cgctggccgc gtctttaaag acagcgacaa attcgatgca aatgattcta tcctaaaaga
     2521 ccaaacacaa gaatggtcag gttcagccac atttacatct gacggaaaaa tccgtttatt
     2581 ctacactgat ttctccggta aacattacgg caaacaaaca ctgacaactg cacaagttaa
     2641 cgtatcagca tcagacagct ctttgaacat caacggtgta gaggattata aatcaatctt
     2701 tgacggtgac ggaaaaacgt atcaaaatgt acagcagttc atcgatgaag gcaactacag
     2761 ctcaggcgac aaccatacgc tgagagatcc tcactacgta gaagataaag gccacaaata
     2821 cttagtattt gaagcaaaca ctggaactga agatggctac caaggcgaag aatctttatt
     2881 taacaaagca tactatggca aaagcacatc attcttccgt caagaaagtc aaaaacttct
     2941 gcaaagcgat aaaaaacgca cggctgagtt agcaaacggc gctctcggta tgattgagct
     3001 aaacgatgat tacacactga aaaaagtgat gaaaccgctg attgcatcta acacagtaac
     3061 agatgaaatt gaacgcgcga acgtctttaa aatgaacggc aaatggtacc tgttcactga
     3121 ctcccgcgga tcaaaaatga cgattgacgg cattacgtct aacgatattt acatgcttgg
     3181 ttatgtttct aattctttaa ctggcccata caagccgctg aacaaaactg gccttgtgtt
     3241 aaaaatggat cttgatccta acgatgtaac ctttacttac tcacacttcg ctgtacctca
     3301 agcgaaagga aacaatgtcg tgattacaag ctatatgaca aacagaggat tctacgcaga
     3361 caaacaatca acgtttgcgc cgagcttcct gctgaacatc aaaggcaaga aaacatctgt
     3421 tgtcaaagac agcatccttg aacaaggaca attaacagtt aacaaataaa aacgcaaaag
     3481 aaaatgccga tgggtaccga gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3541 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3601 cgccctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     3661 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     3721 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     3781 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     3841 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     3901 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     3961 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     4021 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc tgcttccctg
     4081 ctgttttgtg gaatatctac cgactggaaa caggcaaatg caggaaatta ctgaactgag
     4141 gggacaggcg agagacgatg ccaaagagct cctgaaaatc tcgataactc aaaaaatacg
     4201 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     4261 tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt
     4321 atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt
     4381 gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc
     4441 tgtttctatc agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     4501 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     4561 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     4621 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac
     4681 tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagggct
     4741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aaaaggatct
     4801 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5041 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5521 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5581 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5641 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5701 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5761 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5821 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5881 acagctatga catgattacg aattcgagct cggtacccgg ggatcctcta gagtcgacct
     5941 gcaggcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     6001 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     6061 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgat
     6121 aagctagctt cacgc
//
