LOCUS       62056_21390        5826 bp DNA     circular SYN 25-MAR-2019
DEFINITION  Cloning vector pTrcHisA-GFP(tvLAA)-(teF)tvmv, complete sequence.
ACCESSION   MK258729
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5826)
  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 5826)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5826
                     /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)"
     regulatory      368..372
                     /label=RBS
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     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             455..1171
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    1178..1195
                     /label=TVMVp cleave site
                     /note="TVMVp cleave site"
     misc_feature    1202..1234
                     /label=ssrA tag
                     /note="ssrA tag"
     regulatory      1257..1268
                     /label=b0034 RBS
                     /note="b0034 RBS"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1278..1295
                     /label=TEVp cleave site
                     /note="TEVp cleave site"
     misc_feature    1296..1310
                     /label=F degron
                     /note="F degron"
     terminator      2179..2265
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2357..2384
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2404..2495
                     /label=AmpR promoter
     CDS             2496..3353
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3527..4115
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4301..4441)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        4627..4704
                     /label=lacI promoter
     CDS             4705..5784
                     /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 ccatgggcag
      421 cagccatcac catcatcacc acagccagga tcccatggtg agcaagggcg aggagctgtt
      481 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      541 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      601 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      661 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      721 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      781 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      841 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      901 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
      961 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1021 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1081 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1141 gatcactctc ggcatggacg agctgtacaa gggtagcgag accgtgcgct tccagggtag
     1201 cgctgctaac gacgaaaact acgctctggc tgcttaagaa ttcgagctcc gtcgacaaag
     1261 aggagaaagg atctatggaa aacctgtatt ttcagttctt attcgtgcaa gaactgagta
     1321 aagccctgct gaaaggtgtg cgcgatttca acccgattag cgcctgcgtt tgcctgctgg
     1381 aaaatagctc cgacggtcat agcgaacgtc tgtttggcat cggctttggt ccgtacatca
     1441 tcgcgaacca gcacctgttt cgtcgcaaca acggcgaact gaccatcaaa accatgcacg
     1501 gcgagttcaa agtcaaaaac agcacgcagc tgcagatgaa accggttgaa ggtcgcgaca
     1561 tcatcgtgat caaaatggcg aaagacttcc cgccgtttcc gcagaaactg aaattccgtc
     1621 agccgaccat taaagatcgc gtttgcatgg tcagcaccaa cttccagcag aaaagcgtga
     1681 gtagcctggt tagcgaaagt agccacatcg tccacaaaga ggataccagc ttctggcagc
     1741 attggattac gaccaaagac ggtcagtgcg gtagtccgct ggtaagcatc attgacggca
     1801 acattctggg tattcatagc ctgacccata ccaccaacgg cagcaactac ttcgtcgaat
     1861 tcccggaaaa attcgttgcg acctatctgg acgctgccga cggttggtgc aaaaactgga
     1921 aattcaacgc cgacaaaatc tcctggggca gttttaccct ggttgaataa aagcttggct
     1981 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     2041 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     2101 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     2161 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2221 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     2281 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     2341 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     2401 ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2461 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2521 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2581 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2641 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2701 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     2761 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2821 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2881 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2941 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3001 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3061 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3121 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3181 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3241 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3301 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3361 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3421 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3481 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3541 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3601 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3661 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3721 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3781 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3841 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3901 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3961 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4021 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4081 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4141 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4201 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4261 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4321 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     4381 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     4441 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4501 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4561 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt
     4621 tacgttgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     4681 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     4741 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     4801 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     4861 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     4921 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     4981 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     5041 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     5101 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     5161 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     5221 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     5281 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     5341 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     5401 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     5461 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     5521 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
     5581 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     5641 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     5701 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5761 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt
     5821 gatctg
//
