LOCUS       Exported                3101 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMA_Level1D, complete sequence.
ACCESSION   MG252985
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3101)
  AUTHORS   Andreou AI, Nakayama N.
  TITLE     Mobius Assembly: A versatile Golden-Gate framework towards universal
            DNA assembly
  JOURNAL   PLoS ONE 13 (1), e0189892 (2018)
  PUBMED    29293531
REFERENCE   2  (bases 1 to 3101)
  AUTHORS   Andreou AI, Nakayama N.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-OCT-2017) Centre for Synthetic and Systems Biology and
            Institute of Molecular Plant Sciences, University of Edinburgh, Max 
            Born Crescent, Kings Buildings, Edinburgh EH9 3BF, UK
REFERENCE   3  (bases 1 to 3101)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3101
                     /organism="Cloning vector pMA_Level1D"
                     /mol_type="other DNA"
                     /db_xref="taxon:2069627"
     regulatory      2..59
                     /regulatory_class="terminator"
                     /note="his operon terminator"
     terminator      2..59
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     rep_origin      complement(254..842)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(1025..1119)
                     /regulatory_class="terminator"
                     /note="lambda t0 terminator"
     terminator      1025..1119
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1140..1955)
                     /codon_start=1
                     /gene="nptI"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance protein"
                     /label=nptI
                     /protein_id="AUN35406.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(1140..1955)
                     /gene="nptI"
                     /label=nptI
     regulatory      2139..2182
                     /regulatory_class="terminator"
                     /note="bacterial terminator"
     terminator      2139..2182
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2202..2205
                     /label=CTTG overlap
                     /note="CTTG overlap"
     misc_feature    2206..2209
                     /label=GGAG overlap
                     /note="GGAG overlap"
     regulatory      2217..2251
                     /regulatory_class="promoter"
                     /note="Anderson J23106 promoter"
     regulatory      2260..2271
                     /regulatory_class="ribosome_binding_site"
                     /note="B0034 RBS"
     RBS             2260..2271
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2278..2952
                     /codon_start=1
                     /gene="spisPink"
                     /product="spisPink chromoprotein"
                     /label=spisPink
                     /note="chromoprotein from the coral Stylophora pistillata"
                     /protein_id="AUN35407.1"
                     /translation="MSHSKQALADTMKMTWLMEGSVNGHAFTIEGEGTGKPYEGKQSGT
                     FRVTKGGPLPFAFDIVAPTLKYGFKCFMKYPADIPDYFKLAFPEGLTYDRKIAFEDGGC
                     ATATVEMSLKGNTLVHKTNFQGGNFPIDGPVMQKRTLGWEPTSEKMTPCDGIIKGDTIM
                     YLMVEGGKTLKCRYENNYRANKPVLMPPSHFVDLRLTRTNLDKEGLAFKLEEYAVARVL
                     EV"
     gene            2278..2952
                     /gene="spisPink"
                     /label=spisPink
     regulatory      2960..3074
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T7Te double terminator"
     terminator      2960..3031
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3047..3074
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3082..3085
                     /label=CGCT overlap
                     /note="CGCT overlap"