LOCUS       62056_14335        5347 bp DNA     circular SYN 28-MAR-2021
DEFINITION  Cloning vector pLM160-mNeonGreen, complete sequence.
ACCESSION   MT737961
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5347)
  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 5347)
  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..5347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..735
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
     CDS             745..810
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     CDS             1908..2711
                     /codon_start=1
                     /transl_table=11
                     /product="APHVIII"
                     /label=APHVIII
                     /protein_id="QTE34481.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     rep_origin      3084..3629
                     /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(3728..4540)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4990..5292
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
ORIGIN
        1 ggagatctgg gtggctccgg cggccggatg gtgtccaagg gcgaggagga caacatggcg
       61 agcctgcccg cgacgcacga gctgcacatc ttcggcagca tcaacggcgt ggacttcgac
      121 atggtgggcc agggcaccgg caaccccaac gacggctacg aggagctgaa cctgaagtcc
      181 acgaagggcg acctgcagtt cagcccctgg attctggtgc cgcacatcgg ctacggcttc
      241 caccagtacc tgccctaccc ggacggcatg agcccgttcc aggccgcgat ggtggacggc
      301 tcgggctacc aggtgcaccg caccatgcag ttcgaggacg gcgcctccct gaccgtgaac
      361 taccgctaca cgtacgaggg cagccacatc aagggcgagg cccaggtgaa gggcaccggc
      421 ttccccgcgg acggcccggt gatgaccaac tcgctgacgg ccgcggactg gtgccgctcc
      481 aagaagacct accccaacga caagaccatc atcagcacgt tcaagtggtc gtacaccacg
      541 ggcaacggca agcgctaccg ctcgaccgcc cgcaccacgt acacgttcgc gaagcccatg
      601 gccgcgaact acctgaagaa ccagccgatg tacgtgttcc gcaagaccga gctgaagcac
      661 agcaagacgg agctgaactt caaggagtgg cagaaggcgt tcaccgacgt gatgggcatg
      721 gacgagctgt acaagggcgg tggcgactac aaggaccatg acggtgacta taaggatcac
      781 gacatcgact acaaggacga tgacgacaag taagcgttct ggcagcagct ggaccgcctg
      841 taccatggag aagagcttta cttgccggga tggccgattt cgctgattga tacgggatcg
      901 gagctcggag gctttcgcgc taggggctag gcgaagggca gtggtgacca gggtcggtgt
      961 ggggtcggcc cacggtcaat tagccacagg aggatcaggg ggaggtaggc acgtcgactt
     1021 ggtttgcgac cccgcagttt tggcggacgt gctgttgtag atgttagcgt gtgcgtgagc
     1081 cagtggccaa cgtgccacac ccattgagaa gaccaaccaa cttactggca atatctgcca
     1141 atgccatact gcatgtaatg gccaggccat gtgagagttt gccgtgcctg cgcgcgcccc
     1201 gggggcgaag cttggtaccg ctagccttga catgattggt gcgtatgttt gtatgaagct
     1261 acaggactga tttggcgggc tatgagggcg ggggaagctc tggaagggcc gcgatggggc
     1321 gcgcggcgtc cagaaggcgc catacggccc gctggcggca cccatccggt ataaaagccc
     1381 gcgaccccga acggtgacct ccactttcag cgacaaacga gcacttatac atacgcgact
     1441 attctgccgc tatacataac cactcagcta gcttaagatc ccatcaagct tgcatgccgg
     1501 gcgcgccaga aggagcgcag ccaaaccagg atgatgtttg atggggtatt tgagcacttg
     1561 caacccttat ccggaagccc cctggcccac aaaggctagg cgccaatgca agcagttcgc
     1621 atgcagcccc tggagcggtg ccctcctgat aaaccggcca gggggcctat gttctttact
     1681 tttttacaag agaagtcact caacatctta aaatggccag gtgagtcgac gagcaagccc
     1741 ggcggatcag gcagcgtgct tgcagatttg acttgcaacg cccgcattgt gtcgacgaag
     1801 gcttttggct cctctgtcgc tgtctcaagc agcatctaac cctgcgtcgc cgtttccatt
     1861 tgcaggatgg ccactccgcc ctccccggtg ctgaagaatt tcgaagcatg gacgatgcgt
     1921 tgcgtgcact gcggggtcgg tatcccggtt gtgagtgggt tgttgtggag gatggggcct
     1981 cgggggctgg tgtttatcgg cttcggggtg gtgggcggga gttgtttgtc aaggtggcag
     2041 ctctgggggc cggggtgggc ttgttgggtg aggctgagcg gctggtgtgg ttggcggagg
     2101 tggggattcc cgtacctcgt gttgtggagg gtggtgggga cgagagggtc gcctggttgg
     2161 tcaccgaagc ggttccgggg cgtccggcca gtgcgcggtg gccgcgggag cagcggctgg
     2221 acgtggcggt ggcgctcgcg gggctcgctc gttcgctgca cgcgctggac tgggagcggt
     2281 gtccgttcga tcgcagtctc gcggtgacgg tgccgcaggc ggcccgtgct gtcgctgaag
     2341 ggagcgtcga cttggaggat ctggacgagg agcggaaggg gtggtcgggg gagcggcttc
     2401 tcgccgagct ggagcggact cggcctgcgg acgaggatct ggcggtttgc cacggtgacc
     2461 tgtgcccgga caacgtgctg ctcgaccctc gtacctgcga ggtgaccggg ctgatcgacg
     2521 tggggcgggt cggccgtgcg gaccggcact ccgatctcgc gctggtgctg cgcgagctgg
     2581 cccacgagga ggacccgtgg ttcgggccgg agtgttccgc ggcgttcctg cgggagtacg
     2641 ggcgcgggtg ggatggggcg gtatcggagg aaaagctggc gttttaccgg ctgttggacg
     2701 agttcttctg agggacctga tggtgttggt ggctgggtag ggttgcgtcg cgtgggtgac
     2761 agcacagtgt ggacgttggg atccccgctc cgtgtaaatg gaggcgctcg ttgatctgag
     2821 ccttgccccc tgacgaacgg cggtggatgg aagatactgc tctcaagtgc tgaagcggta
     2881 gcttagctcc ccgtttcgtg ctgatcagtc tttttcaaca cgtaaaaagc ggaggagttt
     2941 tgcaattttg ttggttgtaa cgatcctccg ttgattttgg cctctttctc catgggcggg
     3001 ctgggcgtat ttgaagcgta gggataacag ggtaattgca ctgaaatcta gaaatatttt
     3061 atctgattaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct
     3121 gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct
     3181 ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag
     3241 ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc
     3301 cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc
     3361 gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta
     3421 cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac
     3481 cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct
     3541 ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat
     3601 gcttgtcagg ggggcggagc ctatggaaaa accaggtggc acttttcggg gaaatgtgcg
     3661 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     3721 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3781 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3841 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3901 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3961 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     4021 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4081 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4141 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4201 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     4261 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4321 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4381 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     4441 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4501 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4561 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     4621 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cgttaactaa aagccagata
     4681 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     4741 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     4801 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     4861 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     4921 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     4981 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     5041 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     5101 agtgcacgtc tgctgtcaga taaagtcccc cgtgaacttt acccggtggt gcatatcggg
     5161 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     5221 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     5281 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag
     5341 tgttaac
//
