LOCUS       V015947                 7699 bp    DNA     circular SYN 25-MAR-2019
DEFINITION  Exported.
ACCESSION   V015947
VERSION     V015947
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7699)
  AUTHORS   Gao C.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2018) State Key Laboratory of Food Science and
            Technology, Jiangnan University, 1800 Lihu Road, Wu Xi, Jiang Su
            214122, China
REFERENCE   2  (bases 1 to 7699)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (05-DEC-2018) State Key Laboratory of Food Science and Technology,
            Jiangnan University, 1800 Lihu Road, Wu Xi, Jiang Su 214122, China"
FEATURES             Location/Qualifiers
     source          1..7699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        193..222
                     /label="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     misc_feature    263..332
                     /label="rrnG antiterminator"
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             383..406
                     /label="minicistron"
                     /note="synthetic cistron containing a ribosome binding site
                     (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
     CDS             425..442
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             446..478
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             482..505
                     /label="Xpress(TM) tag"
                     /note="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
     misc_feature    521..538
                     /label="TVMVp cleave site"
                     /note="TVMVp cleave site"
     misc_feature    539..553
                     /label="F degron"
                     /note="F degron"
     CDS             560..1303
                     /gene="xylB"
                     /label="D-xylose 1-dehydrogenase"
                     /note="D-xylose 1-dehydrogenase from Caulobacter vibrioides
                     (strain ATCC 19089 / CB15). Accession#: Q9A9Z0"
     misc_feature    1342..1359
                     /label="TEVp cleave site"
                     /note="TEVp cleave site"
     misc_feature    1360..1380
                     /label="F degron"
                     /note="F degron"
     CDS             1381..3105
                     /gene="ptsI"
                     /label="Phosphoenolpyruvate-protein phosphotransferase"
                     /note="Phosphoenolpyruvate-protein phosphotransferase from
                     Escherichia coli (strain K12). Accession#: P08839"
     misc_feature    3144..3161
                     /label="TEVp cleave site"
                     /note="TEVp cleave site"
     misc_feature    3183..3836
                     /label="TVMVp"
                     /note="TVMVp"
     terminator      4052..4138
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4230..4257
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        4277..4368
                     /label="AmpR promoter"
     CDS             4369..5226
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5400..5988
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(6174..6314)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     promoter        6500..6577
                     /label="lacI promoter"
     CDS             6578..7657
                     /label="lacI"
                     /note="lac repressor"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcg atggggatcc gagaccgtgc gcttccagtt
      541 cttattcgtg caagaactga tgtcttctgc tatctacccg tctctgaaag gtaaacgtgt
      601 tgttatcacc ggtggtggtt ctggtatcgg tgctggtctg accgctggtt tcgctcgtca
      661 gggtgctgaa gttatcttcc tggacatcgc tgacgaagac tctcgtgctc tggaagctga
      721 actggctggt tctccgatcc cgccggttta caaacgttgc gacctgatga acctggaagc
      781 tatcaaagct gttttcgctg aaatcggtga cgttgacgtt ctggttaaca acgctggtaa
      841 cgacgaccgt cacaaactgg ctgacgttac cggtgcttac tgggacgaac gtatcaacgt
      901 taacctgcgt cacatgctgt tctgcaccca ggctgttgct ccgggtatga aaaaacgtgg
      961 tggtggtgct gttatcaact tcggttctat ctcttggcac ctgggtctgg aagacctggt
     1021 tctgtacgaa accgctaaag ctggtatcga aggtatgacc cgtgctctgg ctcgtgaact
     1081 gggtccggac gacatccgtg ttacctgcgt tgttccgggt aacgttaaaa ccaaacgtca
     1141 ggaaaaatgg tacaccccgg aaggtgaagc tcagatcgtt gctgctcagt gcctgaaagg
     1201 tcgtatcgtt ccggaaaacg ttgctgctct ggttctgttc ctggcttctg acgacgcttc
     1261 tctgtgcacc ggtcacgaat actggatcga cgctggttgg cgttaagagc tctcacacag
     1321 gacgtcgaca tggcagatct ggaaaatctg tattttcagt ttctgtttgt gcaagaactg
     1381 atgatttcag gcattttagc atccccgggt atcgctttcg gtaaagctct gcttctgaaa
     1441 gaagacgaaa ttgtcattga ccggaaaaaa atttctgccg accaggttga tcaggaagtt
     1501 gaacgttttc tgagcggtcg tgccaaggca tcagcccagc tggaaacgat caaaacgaaa
     1561 gctggtgaaa cgttcggtga agaaaaagaa gccatctttg aagggcatat tatgctgctc
     1621 gaagatgagg agctggagca ggaaatcata gccctgatta aagataagca catgacagct
     1681 gacgcagctg ctcatgaagt tatcgaaggt caggcttctg ccctggaaga gctggatgat
     1741 gaatacctga aagaacgtgc ggctgacgta cgtgatatcg gtaagcgcct gctgcgcaac
     1801 atcctgggcc tgaagattat cgacctgagc gccattcagg atgaagtcat tctggttgcc
     1861 gctgacctga cgccgtccga aaccgcacag ctgaacctga agaaggtgct gggtttcatc
     1921 accgacgcgg gtggccgtac ttcccacacc tctatcatgg cgcgttctct ggaactacct
     1981 gctatcgtgg gtaccggtag cgtcacctct caggtgaaaa atgacgacta tctgattctg
     2041 gatgccgtaa ataatcaggt ttacgtcaat ccaaccaacg aagttattga taaaatgcgc
     2101 gctgttcagg agcaagtggc ttctgaaaaa gcagagcttg ctaaactgaa agatctgcca
     2161 gctattacgc tggacggtca ccaggtagaa gtatgcgcta acattggtac ggttcgtgac
     2221 gttgaaggtg cagagcgtaa cggcgctgaa ggcgttggtc tgtatcgtac tgagttcctg
     2281 ttcatggacc gcgacgcact gcccactgaa gaagaacagt ttgctgctta caaagcagtg
     2341 gctgaagcgt gtggctcgca agcggttatc gttcgtacca tggacatcgg cggcgacaaa
     2401 gagctgccat acatgaactt cccgaaagaa gagaacccgt tcctcggctg gcgcgctatc
     2461 cgtatcgcga tggatcgtag agagatcctg cgcgatcagc tccgcgctat cctgcgtgcc
     2521 tcggctttcg gtaaattgcg cattatgttc ccgatgatca tctctgttga agaagtgcgt
     2581 gcactgcgca aagagatcga aatctacaaa caggaactgc gcgacgaagg taaagcgttt
     2641 gacgagtcaa ttgaaatcgg cgtaatggtg gaaacaccgg ctgccgcaac aattgcacgt
     2701 catttagcca aagaagttga tttctttagt atcggcacca atgatttaac gcagtacact
     2761 ctggcagttg accgtggtaa tgatatgatt tcacaccttt accagccaat gtcaccgtcc
     2821 gtgctgaact tgatcaagca agttattgat gcttctcatg ctgaaggcaa atggactggc
     2881 atgtgtggtg agcttgctgg cgatgaacgt gctacacttc tgttgctggg gatgggtctg
     2941 gacgaattct ctatgagcgc catttctatc ccgcgcatta agaagattat ccgtaacacg
     3001 aacttcgaag atgcgaaggt gttagcagag caggctcttg ctcaaccgac aacggacgag
     3061 ttaatgacgc tggttaacaa gttcattgaa gaaaaaacaa tctgctaaga gctctcacac
     3121 aggacgtcga catggcagat ctggaaaatc tgtattttca gtttctgttt gtgcaagaac
     3181 tgagtaaagc cctgctgaaa ggtgtgcgcg atttcaaccc gattagcgcc tgcgtttgcc
     3241 tgctggaaaa tagctccgac ggtcatagcg aacgtctgtt tggcatcggc tttggtccgt
     3301 acatcatcgc gaaccagcac ctgtttcgtc gcaacaacgg cgaactgacc atcaaaacca
     3361 tgcacggcga gttcaaagtc aaaaacagca cgcagctgca gatgaaaccg gttgaaggtc
     3421 gcgacatcat cgtgatcaaa atggcgaaag acttcccgcc gtttccgcag aaactgaaat
     3481 tccgtcagcc gaccattaaa gatcgcgttt gcatggtcag caccaacttc cagcagaaaa
     3541 gcgtgagtag cctggttagc gaaagtagcc acatcgtcca caaagaggat accagcttct
     3601 ggcagcattg gattacgacc aaagacggtc agtgcggtag tccgctggta agcatcattg
     3661 acggcaacat tctgggtatt catagcctga cccataccac caacggcagc aactacttcg
     3721 tcgaattccc ggaaaaattc gttgcgacct atctggacgc tgccgacggt tggtgcaaaa
     3781 actggaaatt caacgccgac aaaatctcct ggggcagttt taccctggtt gaataagaat
     3841 tcgaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     3901 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     3961 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     4021 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     4081 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     4141 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     4201 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     4261 gtttctacaa actctttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4321 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4381 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4441 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4501 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4561 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     4621 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4681 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4741 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4801 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4861 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4921 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4981 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5041 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5101 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5161 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5221 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5281 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5521 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     6181 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     6241 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     6301 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6361 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6421 ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga
     6481 agcggcatgc atttacgttg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     6541 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     6601 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     6661 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     6721 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     6781 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     6841 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     6901 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     6961 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     7021 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     7081 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     7141 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     7201 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     7261 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     7321 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     7381 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgtcaaccac
     7441 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     7501 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     7561 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7621 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7681 gttagcgcga attgatctg
//
