LOCUS       40924_29856        3101 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMA_Level1D, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3101)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2018"; volume: "13"; issue: "1"; pages: "e0189892"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1025..1119)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1143..1955)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1956..2059)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(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
                     /label=Anderson J23106 promoter
                     /note="Anderson J23106 promoter"
                     /regulatory_class="promoter"
     regulatory      2260..2271
                     /label=B0034 RBS
                     /note="B0034 RBS"
                     /regulatory_class="ribosome_binding_site"
     RBS             2260..2271
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2278..2949
                     /label=spisCP
                     /note="spisCP is a fluorescent protein published in 2008, 
                     derived from Stylophora pistillata. It is reported to be a 
                     tetramer."
     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"
ORIGIN
        1 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
       61 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      241 cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc
     1021 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     1081 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
     1141 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     1201 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     1261 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     1321 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     1381 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     1441 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     1501 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     1561 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     1621 tcttctaata cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca
     1681 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     1741 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     1801 aactctggcg catcgggctt cccatacaat ccatagattg tcgcacctga ttgcccgaca
     1861 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     1921 ctggagcaag acgtttcccg ttgaatatgg ctcattttag cttccttagc tcctgaaaat
     1981 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     2041 cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc taagaaacca ttattatcat
     2101 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     2161 agctttcgct aaggatgatt tctggaattc cacctgcata tcttgggaga gagaccttta
     2221 cggctagctc agtcctaggt atagtgctag ctactagaga aagaggagaa atactaaatg
     2281 tcgcactcaa aacaagcact ggcagatacg atgaaaatga cgtggctgat ggaaggttcg
     2341 gtcaacggtc acgcctttac gattgaaggc gaaggtaccg gcaaaccgta tgaaggtaaa
     2401 cagagtggca cctttcgtgt tacgaaaggc ggtccgctgc cgtttgcgtt tgatattgtc
     2461 gctccgaccc tgaaatacgg tttcaaatgc ttcatgaaat acccggcgga tattccggac
     2521 tactttaaac tggccttccc ggaaggcctg acgtacgatc gtaaaattgc gtttgaagac
     2581 ggcggttgtg cgaccgccac ggtcgaaatg agcctgaagg gtaacaccct ggtgcataaa
     2641 acgaactttc agggcggtaa tttcccgatt gatggtccgg tgatgcaaaa acgcaccctg
     2701 ggctgggaac cgacctctga aaaaatgacg ccgtgcgatg gtattatcaa aggcgacacc
     2761 atcatgtacc tgatggttga aggcggtaaa acgctgaaat gtcgttatga aaacaattac
     2821 cgcgccaaca aaccagtgct gatgccgccg agccactttg tggatctgcg tctgacccgc
     2881 acgaatctgg ataaagaagg tctggcgttc aaactggaag aatatgctgt tgcccgtgtg
     2941 ctggaagtgt aataagcttc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     3001 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     3061 tgggcctttc tgcgggtctc acgctatatg caggtgctgc a
//
