LOCUS       V016421                 6809 bp    DNA     circular SYN 25-MAR-2019
DEFINITION  Exported.
ACCESSION   V016421
VERSION     V016421
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6809)
  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 6809)
  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..6809
                     /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"
     CDS             521..1264
                     /gene="xylB"
                     /label="D-xylose 1-dehydrogenase"
                     /note="D-xylose 1-dehydrogenase from Caulobacter vibrioides
                     (strain ATCC 19089 / CB15). Accession#: Q9A9Z0"
     regulatory      1274..1285
                     /label="b0034 RBS"
                     /note="b0034 RBS"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1304..1321
                     /label="TEVp cleave site"
                     /note="TEVp cleave site"
     misc_feature    1322..1342
                     /label="F degron"
                     /note="F degron"
     CDS             1343..2209
                     /gene="xylC"
                     /label="D-xylonolactone lactonase"
                     /note="D-xylonolactone lactonase from Caulobacter
                     vibrioides (strain ATCC 19089 / CB15). Accession#: Q9A9Z1"
     misc_feature    2219..2230
                     /label="B0034 RBS"
                     /note="B0034 RBS"
     CDS             2237..2950
                     /label="ratiometric pHluorin"
                     /note="pH-sensitive mutant of green fluorescent protein
                     (Miesenbock et al., 1998)"
     terminator      3162..3248
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3340..3367
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3387..3478
                     /label="AmpR promoter"
     CDS             3479..4336
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4510..5098
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(5284..5424)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     promoter        5610..5687
                     /label="lacI promoter"
     CDS             5688..6767
                     /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 atgtcttctg ctatctaccc
      541 gtctctgaaa ggtaaacgtg ttgttatcac cggtggtggt tctggtatcg gtgctggtct
      601 gaccgctggt ttcgctcgtc agggtgctga agttatcttc ctggacatcg ctgacgaaga
      661 ctctcgtgct ctggaagctg aactggctgg ttctccgatc ccgccggttt acaaacgttg
      721 cgacctgatg aacctggaag ctatcaaagc tgttttcgct gaaatcggtg acgttgacgt
      781 tctggttaac aacgctggta acgacgaccg tcacaaactg gctgacgtta ccggtgctta
      841 ctgggacgaa cgtatcaacg ttaacctgcg tcacatgctg ttctgcaccc aggctgttgc
      901 tccgggtatg aaaaaacgtg gtggtggtgc tgttatcaac ttcggttcta tctcttggca
      961 cctgggtctg gaagacctgg ttctgtacga aaccgctaaa gctggtatcg aaggtatgac
     1021 ccgtgctctg gctcgtgaac tgggtccgga cgacatccgt gttacctgcg ttgttccggg
     1081 taacgttaaa accaaacgtc aggaaaaatg gtacaccccg gaaggtgaag ctcagatcgt
     1141 tgctgctcag tgcctgaaag gtcgtatcgt tccggaaaac gttgctgctc tggttctgtt
     1201 cctggcttct gacgacgctt ctctgtgcac cggtcacgaa tactggatcg acgctggttg
     1261 gcgttaagag ctcaaagagg agaaagtcga catggcagat ctggaaaatc tgtattttca
     1321 gtttctgttt gtgcaagaac tgatgaccgc ccaagttact tgtgtttggg atctgaaagc
     1381 caccttaggc gaaggtccta tctggcatgg cgatacttta tggttcgtgg acatcaaaca
     1441 gcgcaaaatc cacaattatc acccggccac tggtgagcgt ttcagtttcg atgcaccgga
     1501 ccaagttacc tttttagccc cgattgtggg cgccaccggc tttgtggtgg gtctgaaaac
     1561 cggcatccac cgttttcatc cggcaaccgg cttcagctta ctgctggaag tggaagatgc
     1621 cgcactgaac aaccgcccga acgacgccac agtggatgca caaggtcgtt tatggtttgg
     1681 caccatgcac gatggtgagg aaaacaatag cggctcttta tatcgcatgg atctgaccgg
     1741 tgtggcacgc atggaccgtg acatttgcat taccaacggc ccgtgtgtga gcccggacgg
     1801 caaaaccttt taccacaccg acactttaga gaagaccatc tacgccttcg atctggcaga
     1861 agacggttta ctgagtaaca agcgcgtgtt cgttcagttt gctttaggtg atgatgtgta
     1921 tccggacggc agcgtggtgg atagtgaagg ctatctgtgg accgcactgt ggggcggttt
     1981 cggtgcagtg cgctttagcc cgcaaggtga tgccgttacc cgcattgaac tgccggcacc
     2041 gaacgtgaca aaaccgtgct ttggtggtcc ggatttaaaa actttatact tcaccaccgc
     2101 ccgtaaaggt ttaagtgacg aaactttagc acaatacccg ctggctggtg gcgtttttgc
     2161 agttcccgtt gatgtggccg gccaaccgca gcatgaagtg cgtctggttt aagaattcaa
     2221 agaggagaaa tctagaatga gtaaaggaga agaacttttc actggagttg tcccaattct
     2281 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
     2341 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt
     2401 tccatggcca acacttgtca ctactttctc ttatggtgtt caatgctttt caagataccc
     2461 agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     2521 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga
     2581 aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag atgatggaaa
     2641 cattcttgga cacaaattgg aatacaacta taacgagcac ttggtgtaca tcatggcaga
     2701 caaacaaaag aatggtacca aagctatctt tcaagttcac cacaacattg aagatggagg
     2761 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     2821 accagacaac cattacctgc acacacaatc tgccctttcg aaagatccca acgaaaagag
     2881 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga
     2941 actatacaaa taaaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
     3001 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     3061 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     3121 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     3181 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     3241 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     3301 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
     3361 gcctttttgc gtttctacaa actctttttg tttatttttc taaatacatt caaatatgta
     3421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3661 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3721 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3781 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4021 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4081 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4321 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4621 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4861 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4921 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4981 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5041 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5101 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5161 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5221 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5281 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     5401 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     5461 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5521 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     5581 gcgaaggcga agcggcatgc atttacgttg acaccatcga atggcgcaaa acctttcgcg
     5641 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     5701 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     5761 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     5821 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     5881 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     5941 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     6001 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     6061 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     6121 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     6181 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     6241 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     6301 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     6361 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     6421 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     6481 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     6541 cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     6601 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     6661 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6721 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6781 attaatgtga gttagcgcga attgatctg
//
