LOCUS       62056_14330        5356 bp DNA     circular SYN 28-MAR-2021
DEFINITION  Cloning vector pLM099-Venus, complete sequence.
ACCESSION   MT737960
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   Emrich-Mills TZ, Yates G, Barrett J, Girr P, Grouneva I, Lau CS, 
            Walker CE, Kwok TK, Davey JW, Johnson MP, Mackinder LCM.
  TITLE     A recombineering pipeline to clone large and complex genes in 
            Chlamydomonas
  JOURNAL   Plant Cell (2021) In press
  PUBMED    33723601
REFERENCE   2  (bases 1 to 5356)
  AUTHORS   Emrich-Mills TZ, Yates G, Barrett J, Grouneva I, Lau CS, Walker CE, 
            Kwok TK, Davey JW, Johnson MP, Mackinder LCM.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUL-2020) Molecular Biology and Biotechnology, 
            University of Sheffield, Western Bank, Sheffield, South Yorkshire 
            S10 2TN, United Kingdom
REFERENCE   3  (bases 1 to 5356)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell 
            (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUL-2020) Molecular Biology and Biotechnology, University of 
            Sheffield, Western Bank, Sheffield, South Yorkshire S10 2TN, United 
            Kingdom"
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..744
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
     CDS             796..819
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1917..2720
                     /codon_start=1
                     /transl_table=11
                     /product="APHVIII"
                     /label=APHVIII
                     /note="paromomycin resistance marker"
                     /protein_id="QTE34478.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     rep_origin      3093..3638
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(3737..4549)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4999..5301
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
ORIGIN
        1 ggagatctgg gtggctccgg cggccggatg gtgagcaagg gcgaggagct gttcaccggc
       61 gtggtgccca tcctggtgga gctggacggc gacgtgaacg gccacaagtt cagcgtgagc
      121 ggcgagggcg agggcgacgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
      181 ggcaagctgc ccgtgccctg gcccaccctg gtgaccaccc tgggctacgg cctgcagtgc
      241 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagagcgc catgcccgag
      301 ggctacgtgc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      361 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      421 aaggaggacg gcaacatcct gggccacaag ctggagtaca actacaacag ccacaacgtg
      481 tacatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
      541 atcgaggacg gcggcgtgca gctggccgac cactaccagc agaacacccc catcggcgac
      601 ggccccgtgc tgctgcccga caaccactac ctgagctacc agagcgccct gagcaaggac
      661 cccaacgaga agcgcgacca catggtgctg ctggagttcg tgaccgccgc cggcatcacc
      721 ctgggcatgg acgagctgat caagggcggt ggcgactaca aggaccatga cggtgactat
      781 aaggatcacg acatcgacta caaggacgat gacgacaagt aagcgttctg gcagcagctg
      841 gaccgcctgt accatggaga agagctttac ttgccgggat ggccgatttc gctgattgat
      901 acgggatcgg agctcggagg ctttcgcgct aggggctagg cgaagggcag tggtgaccag
      961 ggtcggtgtg gggtcggccc acggtcaatt agccacagga ggatcagggg gaggtaggca
     1021 cgtcgacttg gtttgcgacc ccgcagtttt ggcggacgtg ctgttgtaga tgttagcgtg
     1081 tgcgtgagcc agtggccaac gtgccacacc cattgagaag accaaccaac ttactggcaa
     1141 tatctgccaa tgccatactg catgtaatgg ccaggccatg tgagagtttg ccgtgcctgc
     1201 gcgcgccccg ggggcgaagc ttggtaccgc tagccttgac atgattggtg cgtatgtttg
     1261 tatgaagcta caggactgat ttggcgggct atgagggcgg gggaagctct ggaagggccg
     1321 cgatggggcg cgcggcgtcc agaaggcgcc atacggcccg ctggcggcac ccatccggta
     1381 taaaagcccg cgaccccgaa cggtgacctc cactttcagc gacaaacgag cacttataca
     1441 tacgcgacta ttctgccgct atacataacc actcagctag cttaagatcc catcaagctt
     1501 gcatgccggg cgcgccagaa ggagcgcagc caaaccagga tgatgtttga tggggtattt
     1561 gagcacttgc aacccttatc cggaagcccc ctggcccaca aaggctaggc gccaatgcaa
     1621 gcagttcgca tgcagcccct ggagcggtgc cctcctgata aaccggccag ggggcctatg
     1681 ttctttactt ttttacaaga gaagtcactc aacatcttaa aatggccagg tgagtcgacg
     1741 agcaagcccg gcggatcagg cagcgtgctt gcagatttga cttgcaacgc ccgcattgtg
     1801 tcgacgaagg cttttggctc ctctgtcgct gtctcaagca gcatctaacc ctgcgtcgcc
     1861 gtttccattt gcaggatggc cactccgccc tccccggtgc tgaagaattt cgaagcatgg
     1921 acgatgcgtt gcgtgcactg cggggtcggt atcccggttg tgagtgggtt gttgtggagg
     1981 atggggcctc gggggctggt gtttatcggc ttcggggtgg tgggcgggag ttgtttgtca
     2041 aggtggcagc tctgggggcc ggggtgggct tgttgggtga ggctgagcgg ctggtgtggt
     2101 tggcggaggt ggggattccc gtacctcgtg ttgtggaggg tggtggggac gagagggtcg
     2161 cctggttggt caccgaagcg gttccggggc gtccggccag tgcgcggtgg ccgcgggagc
     2221 agcggctgga cgtggcggtg gcgctcgcgg ggctcgctcg ttcgctgcac gcgctggact
     2281 gggagcggtg tccgttcgat cgcagtctcg cggtgacggt gccgcaggcg gcccgtgctg
     2341 tcgctgaagg gagcgtcgac ttggaggatc tggacgagga gcggaagggg tggtcggggg
     2401 agcggcttct cgccgagctg gagcggactc ggcctgcgga cgaggatctg gcggtttgcc
     2461 acggtgacct gtgcccggac aacgtgctgc tcgaccctcg tacctgcgag gtgaccgggc
     2521 tgatcgacgt ggggcgggtc ggccgtgcgg accggcactc cgatctcgcg ctggtgctgc
     2581 gcgagctggc ccacgaggag gacccgtggt tcgggccgga gtgttccgcg gcgttcctgc
     2641 gggagtacgg gcgcgggtgg gatggggcgg tatcggagga aaagctggcg ttttaccggc
     2701 tgttggacga gttcttctga gggacctgat ggtgttggtg gctgggtagg gttgcgtcgc
     2761 gtgggtgaca gcacagtgtg gacgttggga tccccgctcc gtgtaaatgg aggcgctcgt
     2821 tgatctgagc cttgccccct gacgaacggc ggtggatgga agatactgct ctcaagtgct
     2881 gaagcggtag cttagctccc cgtttcgtgc tgatcagtct ttttcaacac gtaaaaagcg
     2941 gaggagtttt gcaattttgt tggttgtaac gatcctccgt tgattttggc ctctttctcc
     3001 atgggcgggc tgggcgtatt tgaagcgtag ggataacagg gtaattgcac tgaaatctag
     3061 aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc
     3121 tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct
     3181 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     3241 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     3301 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     3361 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     3421 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     3481 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     3541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     3601 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa ccaggtggca cttttcgggg
     3661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3721 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     3781 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     3841 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     3901 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     3961 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     4021 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     4081 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     4141 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     4201 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     4261 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     4321 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     4381 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     4441 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     4501 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
     4561 tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag
     4621 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc gttaactaaa
     4681 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     4741 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     4801 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     4861 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     4921 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     4981 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     5041 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     5101 cccctggcca gtgcacgtct gctgtcagat aaagtccccc gtgaacttta cccggtggtg
     5161 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     5221 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     5281 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     5341 cgaccatagt gttaac
//
