LOCUS       62056_14340        5277 bp DNA     circular SYN 28-MAR-2021
DEFINITION  Cloning vector pLM161-Venus, complete sequence.
ACCESSION   MT737962
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  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   2  (bases 1 to 5277)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (08-JUL-2020) Molecular Biology and Biotechnology, University of 
            Sheffield, Western Bank, Sheffield, South Yorkshire S10 2TN, United 
            Kingdom"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5277
                     /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             join(1562..1648,1794..2705)
                     /codon_start=1
                     /transl_table=11
                     /product="APHVII"
                     /label=APHVII
                     /note="hygromycin resistance marker"
                     /protein_id="QTE34484.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPGPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     rep_origin      3014..3559
                     /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(3658..4470)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4920..5222
                     /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 tagcttcgat atcaagcttc tttcttgcgc
     1261 tatgacactt ccagcaaaag gtagggcggg ctgcgagacg gcttcccggc gctgcatgca
     1321 acaccgatga tgcttcgacc ccccgaagct ccttcggggc tgcatgggcg ctccgatgcc
     1381 gctccagggc gagcgctgtt taaatagcca ggcccccgat tgcaaagaca ttatagcgag
     1441 ctaccaaagc catattcaaa cacctagatc actaccactt ctacacaggc cactcgagct
     1501 tgtgatcgca ctccgctaag ggggcgcctc ttcctcttcg tttcagtcac aacccgcaaa
     1561 catgacacaa gaatccctgt tacttctcga ccgtattgat tcggatgatt cctacgcgag
     1621 cctgcggaac gaccaggaat tctgggaggt gagtcgacga gcaagcccgg cggatcaggc
     1681 agcgtgcttg cagatttgac ttgcaacgcc cgcattgtgt cgacgaaggc ttttggctcc
     1741 tctgtcgctg tctcaagcag catctaaccc tgcgtcgccg tttccatttg cagccgctgg
     1801 cccgccgagc cctggaggag ctcgggctgc cggtgccgcc ggtgctgcgg gtgcccggcg
     1861 agagcaccaa ccccgtactg gtcggcgagc ccggcccggt gatcaagctg ttcggcgagc
     1921 actggtgcgg tccggagagc ctcgcgtcgg agtcggaggc gtacgcggtc ctggcggacg
     1981 ccccggtgcc ggtgccccgc ctcctcggcc gcggcgagct gcggcccggc accggagcct
     2041 ggccgtggcc ctacctggtg atgagccgga tgaccggcac cacctggcgg tccgcgatgg
     2101 acggcacgac cgaccggaac gcgctgctcg ccctggcccg cgaactcggc cgggtgctcg
     2161 gccggctgca cagggtgccg ctgaccggga acaccgtgct caccccccat tccgaggtct
     2221 tcccggaact gctgcgggaa cgccgcgcgg cgaccgtcga ggaccaccgc gggtggggct
     2281 acctctcgcc ccggctgctg gaccgcctgg aggactggct gccggacgtg gacacgctgc
     2341 tggccggccg cgaaccccgg ttcgtccacg gcgacctgca cgggaccaac atcttcgtgg
     2401 acctggccgc gaccgaggtc accgggatcg tcgacttcac cgacgtctat gcgggagact
     2461 cccgctacag cctggtgcaa ctgcatctca acgccttccg gggcgaccgc gagatcctgg
     2521 ccgcgctgct cgacggggcg cagtggaagc ggaccgagga cttcgcccgc gaactgctcg
     2581 ccttcacctt cctgcacgac ttcgaggtgt tcgaggagac cccgctggat ctctccggct
     2641 tcaccgatcc ggaggaactg gcgcagttcc tctgggggcc gccggacacc gcccccggcg
     2701 cctgataagg atccccgctc cgtgtaaatg gaggcgctcg ttgatctgag ccttgccccc
     2761 tgacgaacgg cggtggatgg aagatactgc tctcaagtgc tgaagcggta gcttagctcc
     2821 ccgtttcgtg ctgatcagtc tttttcaaca cgtaaaaagc ggaggagttt tgcaattttg
     2881 ttggttgtaa cgatcctccg ttgattttgg cctctttctc catgggcggg ctgggcgtat
     2941 ttgaagcgta gggataacag ggtaattgca ctgaaatcta gaaatatttt atctgattaa
     3001 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     3061 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     3121 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     3181 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     3241 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3301 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     3361 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     3421 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     3481 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     3541 ggggcggagc ctatggaaaa accaggtggc acttttcggg gaaatgtgcg cggaacccct
     3601 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     3661 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     3721 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     3781 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     3841 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     3901 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     3961 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     4021 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     4081 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     4141 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     4201 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     4261 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     4321 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     4381 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     4441 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     4501 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4561 cagaccccgt agaaaagatc aaaggatctt cgttaactaa aagccagata acagtatgcg
     4621 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     4681 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     4741 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     4801 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     4861 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     4921 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     4981 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     5041 tgctgtcaga taaagtcccc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     5101 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     5161 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     5221 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgttaac
//
