LOCUS       V015983                 6446 bp    DNA     circular SYN 25-MAR-2019
DEFINITION  Exported.
ACCESSION   V015983
VERSION     V015983
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6446)
  AUTHORS   Gao C.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-NOV-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 6446)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (28-NOV-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..6446
                     /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             482..505
                     /label="Xpress(TM) tag"
                     /note="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
     CDS             521..1801
                     /gene="gltA"
                     /label="Citrate synthase"
                     /note="Citrate synthase from Escherichia coli (strain K12).
                     Accession#: P0ABH7"
     misc_feature    1808..1825
                     /label="tev protease site"
                     /note="tev protease site"
     misc_feature    1826..1840
                     /label="F-degron"
                     /note="F-degron"
     CDS             1847..1873
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1874..2587
                     /label="Venus"
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
     terminator      2799..2885
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2977..3004
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3024..3115
                     /label="AmpR promoter"
     CDS             3116..3973
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4147..4735
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(4921..5061)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     promoter        5247..5324
                     /label="lacI promoter"
     CDS             5325..6404
                     /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 atggctgata caaaagcaaa
      541 actcaccctc aacggggata cagctgttga actggatgtg ctgaaaggca cgctgggtca
      601 agatgttatt gatatccgta ctctcggttc aaaaggtgtg ttcacctttg acccaggctt
      661 cacttcaacc gcatcctgcg aatctaaaat tacttttatt gatggtgatg aaggtatttt
      721 gctgcaccgc ggtttcccga tcgatcagct ggcgaccgat tctaactacc tggaagtttg
      781 ttacatcctg ctgaatggtg aaaaaccgac tcaggaacag tatgacgaat ttaaaactac
      841 ggtgacccgt cataccatga tccacgagca gattacccgt ctgttccatg ctttccgtcg
      901 cgactcgcat ccaatggcag tcatgtgtgg tattaccggc gcgctggcgg cgttctatca
      961 cgactcgctg gatgttaaca atcctcgtca ccgtgaaatt gccgcgttcc gcctgctgtc
     1021 gaaaatgccg accatggccg cgatgtgtta caagtattcc attggtcagc catttgttta
     1081 cccgcgcaac gatctctcct acgccggtaa cttcctgaat atgatgttct ccacgccgtg
     1141 cgaaccgtat gaagttaatc cgattctgga acgtgctatg gaccgtattc tgatcctgca
     1201 cgctgaccat gaacagaacg cctctacctc caccgtgcgt accgctggct cttcgggtgc
     1261 gaacccgttt gcctgtatcg cagcaggtat tgcttcactg tggggacctg cgcacggcgg
     1321 tgctaacgaa gcggcgctga aaatgctgga agaaatcagc tccgttaaac acattccgga
     1381 atttgttcgt cgtgcgaaag acaaaaatga ttctttccgc ctgatgggct tcggtcaccg
     1441 cgtgtacaaa aattacgacc cgcgcgccac cgtaatgcgt gaaacctgcc atgaagtgct
     1501 gaaagagctg ggcacgaagg atgacctgct ggaagtggct atggagctgg aaaacatcgc
     1561 gctgaacgac ccgtacttta tcgagaagaa actgtacccg aacgtcgatt tctactctgg
     1621 tatcatcctg aaagcgatgg gtattccgtc ttccatgttc accgtcattt tcgcaatggc
     1681 acgtaccgtt ggctggatcg cccactggag cgaaatgcac agtgacggta tgaagattgc
     1741 ccgtccgcgt cagctgtata caggatatga aaaacgcgac tttaaaagcg atatcaagcg
     1801 tggatccgaa aacctgtatt ttcagttctt attcgtgcaa ggtagctacc cgtacgacgt
     1861 tccggactac gctgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1921 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1981 tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc
     2041 ctggcccacc ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc gctaccccga
     2101 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2161 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2221 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2281 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa
     2341 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
     2401 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2461 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2521 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2581 gtacaagtaa aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     2641 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     2701 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     2761 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     2821 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     2881 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     2941 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     3001 tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa atatgtatcc
     3061 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3301 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     3361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3421 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3661 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3721 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3961 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4261 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     4321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4921 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     4981 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     5041 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5101 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5161 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg
     5221 aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gcgcaaaacc tttcgcggta
     5281 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     5341 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5401 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5461 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5521 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5581 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5641 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5701 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5761 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5821 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5881 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     5941 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     6001 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     6061 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     6121 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     6181 caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     6241 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     6301 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6361 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6421 aatgtgagtt agcgcgaatt gatctg
//
