LOCUS       62056_21385        5958 bp DNA     circular SYN 25-MAR-2019
DEFINITION  Cloning vector pTrcHisA-GFP(teLAA)-(teF)MCP, complete sequence.
ACCESSION   MK258730
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  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 5958)
  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..5958
                     /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..1237
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             1244..1264
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    1268..1300
                     /label=ssrA tag
                     /note="ssrA tag"
     regulatory      1310..1321
                     /label=b0034 RBS
                     /note="b0034 RBS"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1337..1354
                     /label=TEVp cleave site
                     /note="TEVp cleave site"
     misc_feature    1355..1369
                     /label=F degron
                     /note="F degron"
     CDS             1388..2080
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     terminator      2311..2397
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2489..2516
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2536..2627
                     /label=AmpR promoter
     CDS             2628..3485
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3659..4247
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4433..4573)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        4759..4836
                     /label=lacI promoter
     CDS             4837..5916
                     /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 atggtgagca agggcgagga
      541 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      601 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      661 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      721 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      781 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      841 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      901 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      961 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1021 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1081 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1141 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1201 cgccgggatc actctcggca tggacgagct gtacaagggt agcgaaaacc tgtattttca
     1261 gggtagcgct gctaacgacg aaaactacgc tctggctgct taaagatcta aagaggagaa
     1321 aggatctatg ggtagcgaaa acctgtattt tcagttctta ttcgtgcaag aactgatggt
     1381 gagcaagatg tcagaattaa ttaaagaaaa tatgcacatg aaattatata tggaaggtac
     1441 tgtcaacaat catcatttca aatgcacatc cgaaggtgaa ggtaaaccat atgaaggcac
     1501 acaaacaatg cgcatcaaag cagttgaagg tggacccctg ccctttgcgt ttgacattct
     1561 cgcaacgagc tttatgtacg ggtctaaaac ttttatcaat cacacccaag gcattcctga
     1621 cttttttaaa cagtcctttc ctgaaggctt tacctgggaa cgtgtaacaa cttatgaaga
     1681 tggcggtgta cttacagcaa ctcaagatac gagtttacaa gatggctgtc tgatttacaa
     1741 tgttaaaatc cgtggcgtaa atttcccgag taacggaccc gtaatgcaaa aaaaaactct
     1801 tggttgggaa gcatcaacag aaaccttata tcctgcggac ggtggcttag aaggacgcgc
     1861 agacatggca ctgaaattag ttggaggcgg tcatttaatc tgcaacctga aaacaaccta
     1921 tcgttccaaa aaacccgcta aaaaccttaa aatgcctgga gtatactatg ttgatcgtcg
     1981 cttagaacgt attaaagaag ctgataaaga aacctacgtt gaacaacatg aagtagccgt
     2041 agcccgttat tgtgaccttc cgtcgaaatt aggacatcgt tgaggtacca tatgggaatt
     2101 cgaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     2161 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     2221 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     2281 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     2341 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     2401 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     2461 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     2521 tttctacaaa ctctttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2581 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2641 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2701 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2761 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2821 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2881 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2941 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3001 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3061 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3121 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3181 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3241 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3301 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3361 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3421 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3481 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3541 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3601 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3661 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3721 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3781 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3841 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3901 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3961 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4021 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4081 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4141 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4201 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4261 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4321 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4381 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4441 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     4501 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4561 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4621 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4681 gttttcaccg tcatcaccga aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa
     4741 gcggcatgca tttacgttga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     4801 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     4861 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     4921 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     4981 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     5041 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     5101 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     5161 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     5221 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     5281 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     5341 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     5401 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     5461 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     5521 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     5581 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     5641 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gtcaaccacc
     5701 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     5761 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     5821 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5881 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     5941 ttagcgcgaa ttgatctg
//
