LOCUS       40924_37073        3430 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRI857, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3430)
  AUTHORS   Zhang K, Su H, Yang M, Ge J, Li G, Yi J, Wang Y.
  TITLE     Construction of a thermo-sensitive pRI857 vector for efficient DNA 
            capturing in Escherichia coli
  JOURNAL   Biotechnol. Lett. (2017) In press
  PUBMED    28251389
REFERENCE   2  (bases 1 to 3430)
  AUTHORS   Zhang K, Wang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-AUG-2016) College of Environmental and Biological 
            Engineering, Nanjing University of Science and Technology, 200 
            Xiaolingwei, Nanjing, Jiangsu 210094, China
REFERENCE   3  (bases 1 to 3430)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3430)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Lett. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-AUG-2016) College of Environmental and Biological Engineering, 
            Nanjing University of Science and Technology, 200 Xiaolingwei, 
            Nanjing, Jiangsu 210094, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..795)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(797..837)
                     /gene="KanR"
                     /regulatory_class="promoter"
     RBS             799..807
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    950..971
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        986..1016
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1024..1040
                     /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)."
     CDS             1060..1770
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     regulatory      2026..2062
                     /regulatory_class="terminator"
     primer_bind     complement(2117..2133)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2162..2180)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(2204..2237)
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2259..2275)
                     /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)."
     promoter        complement(2283..2313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2328..2349)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2637..3225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
       61 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
      121 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
      181 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
      241 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
      301 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
      361 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
      421 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
      481 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
      541 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
      601 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
      661 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
      721 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
      781 atcttgttca atcatacaac ctccttagta catgcaacca ttatcaccgc cagaggtaaa
      841 atagtcaaca cgcacggtgt tagatattta tcccttgcgg tgatagattt aacgtcagct
      901 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      961 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1021 gaattgtgag cggataacaa tttcacacag gaaacagcta tgagcacaaa aaagaaacca
     1081 ttaacacaag agcagcttga ggacgcacgt cgccttaaag caatttatga aaaaaagaaa
     1141 aatgaacttg gcttatccca ggaatctgtc gcagacaaga tggggatggg gcagtcaggc
     1201 gttggtgctt tatttaatgg catcaatgca ttaaatgctt ataacgccgc attgcttaca
     1261 aaaattctca aagttagcgt tgaagaattt agcccttcaa tcgccagaga aatctacgag
     1321 atgtatgaag cggttagtat gcagccgtca cttagaagtg agtatgagta ccctgttttt
     1381 tctcatgttc aggcagggat gttctcacct aagcttagaa cctttaccaa aggtgatgcg
     1441 gagagatggg taagcacaac caaaaaagcc agtgattctg cattctggct tgaggttgaa
     1501 ggtaattcca tgaccgcacc aacaggctcc aagccaagct ttcctgacgg aatgttaatt
     1561 ctcgttgacc ctgagcaggc tgttgagcca ggtgatttct gcatagccag acttgggggt
     1621 gatgagttta ccttcaagaa actgatcagg gatagcggtc aggtgttttt acaaccacta
     1681 aacccacagt acccaatgat cccatgcaat gagagttgtt ccgttgtggg gaaagttatc
     1741 gctagtcagt ggcctgaaga gacgtttggc tgatcggcaa ggtgttctgg ttaactgatg
     1801 aactatggga ccaaataaaa tatattgaag aaattaattg attacagaaa ctccttaaaa
     1861 ggagtttttt agttacctta attgaaataa acgaaataaa aactcgcaat taagcgagtt
     1921 ggaatttaaa tatgatatct acattatcag cagtaacatc aacctttgat acaaggttgt
     1981 tgcatttacc ttgattgaga tgttaattgt gttggcaatt atcagaaaaa agaacccgag
     2041 tgggttcttt tttatttttt aatctagcta gaactagtgg atcccccggg ctgcaggaat
     2101 tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc agcttttgtt
     2161 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtctaccgtt cgtatagcat
     2221 acattatacg aagttatata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3061 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3421 cctatctcag
//
