LOCUS       62056_21395        5129 bp DNA     circular SYN 25-MAR-2019
DEFINITION  Cloning vector pTrcHisA-GFP-(te)-(LAA), complete sequence.
ACCESSION   MK238519
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5129)
  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 5129)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5129
                     /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"
     regulatory      473..484
                     /label=B0034
                     /note="B0034"
                     /regulatory_class="ribosome_binding_site"
     CDS             491..1207
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             1214..1234
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    1238..1270
                     /label=ssrA tag
                     /note="ssrA tag"
     terminator      1482..1568
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1660..1687
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1707..1798
                     /label=AmpR promoter
     CDS             1799..2656
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2830..3418
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3604..3744)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        3930..4007
                     /label=lacI promoter
     CDS             4008..5087
                     /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 tccgaattcg agctccgtcg acaaagagga
      481 gaaaggatct atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      541 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      601 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      661 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      721 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      781 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      841 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      901 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      961 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1021 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1081 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1141 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1201 gtacaagggt agcgaaaacc tgtattttca gggtagcgct gctaacgacg aaaactacgc
     1261 tctggctgct taaaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
     1321 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     1381 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     1441 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     1501 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1561 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     1621 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
     1681 gcctttttgc gtttctacaa actctttttg tttatttttc taaatacatt caaatatgta
     1741 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1801 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1861 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1921 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1981 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2041 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2101 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2161 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2221 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2281 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2341 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2401 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2461 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2521 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2581 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2641 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2701 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2761 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2821 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2881 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2941 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3001 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3061 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3121 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3181 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3241 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3301 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3361 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3421 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3481 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3541 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3601 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     3661 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3721 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3781 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3841 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     3901 gcgaaggcga agcggcatgc atttacgttg acaccatcga atggcgcaaa acctttcgcg
     3961 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     4021 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     4081 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     4141 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     4201 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     4261 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     4321 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     4381 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     4441 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     4501 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     4561 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     4621 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     4681 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     4741 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     4801 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     4861 cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     4921 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     4981 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5041 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5101 attaatgtga gttagcgcga attgatctg
//
