LOCUS       62056_14345        5256 bp DNA     circular SYN 28-MAR-2021
DEFINITION  Cloning vector pLM162-mScarlet-i, complete sequence.
ACCESSION   MT737963
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  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 5256)
  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..5256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..723
                     /label=mScarlet-I
                     /note="bright monomeric red fluorescent protein evolved
                     from a synthetic template (Bindels et al., 2016)"
     CDS             775..798
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             join(1541..1627,1773..2684)
                     /codon_start=1
                     /transl_table=11
                     /product="APHVII"
                     /label=APHVII
                     /note="hygromycin resistance marker"
                     /protein_id="QTE34487.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPGPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     rep_origin      2993..3538
                     /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(3637..4449)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4899..5201
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
ORIGIN
        1 ggagatctgg gtggctccgg cggccggatg gtgagcaagg gcgaggcagt gatcaaggag
       61 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
      121 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
      181 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
      241 gccttcatca agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc
      301 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac
      361 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
      421 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg
      481 taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc
      541 ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc
      601 ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg
      661 gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga cgagctgtac
      721 aagggcggtg gcgactacaa ggaccatgac ggtgactata aggatcacga catcgactac
      781 aaggacgatg acgacaagta agcgttctgg cagcagctgg accgcctgta ccatggagaa
      841 gagctttact tgccgggatg gccgatttcg ctgattgata cgggatcgga gctcggaggc
      901 tttcgcgcta ggggctaggc gaagggcagt ggtgaccagg gtcggtgtgg ggtcggccca
      961 cggtcaatta gccacaggag gatcaggggg aggtaggcac gtcgacttgg tttgcgaccc
     1021 cgcagttttg gcggacgtgc tgttgtagat gttagcgtgt gcgtgagcca gtggccaacg
     1081 tgccacaccc attgagaaga ccaaccaact tactggcaat atctgccaat gccatactgc
     1141 atgtaatggc caggccatgt gagagtttgc cgtgcctgcg cgcgccccgg gggcgaagct
     1201 tggtaccgct agcttcgata tcaagcttct ttcttgcgct atgacacttc cagcaaaagg
     1261 tagggcgggc tgcgagacgg cttcccggcg ctgcatgcaa caccgatgat gcttcgaccc
     1321 cccgaagctc cttcggggct gcatgggcgc tccgatgccg ctccagggcg agcgctgttt
     1381 aaatagccag gcccccgatt gcaaagacat tatagcgagc taccaaagcc atattcaaac
     1441 acctagatca ctaccacttc tacacaggcc actcgagctt gtgatcgcac tccgctaagg
     1501 gggcgcctct tcctcttcgt ttcagtcaca acccgcaaac atgacacaag aatccctgtt
     1561 acttctcgac cgtattgatt cggatgattc ctacgcgagc ctgcggaacg accaggaatt
     1621 ctgggaggtg agtcgacgag caagcccggc ggatcaggca gcgtgcttgc agatttgact
     1681 tgcaacgccc gcattgtgtc gacgaaggct tttggctcct ctgtcgctgt ctcaagcagc
     1741 atctaaccct gcgtcgccgt ttccatttgc agccgctggc ccgccgagcc ctggaggagc
     1801 tcgggctgcc ggtgccgccg gtgctgcggg tgcccggcga gagcaccaac cccgtactgg
     1861 tcggcgagcc cggcccggtg atcaagctgt tcggcgagca ctggtgcggt ccggagagcc
     1921 tcgcgtcgga gtcggaggcg tacgcggtcc tggcggacgc cccggtgccg gtgccccgcc
     1981 tcctcggccg cggcgagctg cggcccggca ccggagcctg gccgtggccc tacctggtga
     2041 tgagccggat gaccggcacc acctggcggt ccgcgatgga cggcacgacc gaccggaacg
     2101 cgctgctcgc cctggcccgc gaactcggcc gggtgctcgg ccggctgcac agggtgccgc
     2161 tgaccgggaa caccgtgctc accccccatt ccgaggtctt cccggaactg ctgcgggaac
     2221 gccgcgcggc gaccgtcgag gaccaccgcg ggtggggcta cctctcgccc cggctgctgg
     2281 accgcctgga ggactggctg ccggacgtgg acacgctgct ggccggccgc gaaccccggt
     2341 tcgtccacgg cgacctgcac gggaccaaca tcttcgtgga cctggccgcg accgaggtca
     2401 ccgggatcgt cgacttcacc gacgtctatg cgggagactc ccgctacagc ctggtgcaac
     2461 tgcatctcaa cgccttccgg ggcgaccgcg agatcctggc cgcgctgctc gacggggcgc
     2521 agtggaagcg gaccgaggac ttcgcccgcg aactgctcgc cttcaccttc ctgcacgact
     2581 tcgaggtgtt cgaggagacc ccgctggatc tctccggctt caccgatccg gaggaactgg
     2641 cgcagttcct ctgggggccg ccggacaccg cccccggcgc ctgataagga tccccgctcc
     2701 gtgtaaatgg aggcgctcgt tgatctgagc cttgccccct gacgaacggc ggtggatgga
     2761 agatactgct ctcaagtgct gaagcggtag cttagctccc cgtttcgtgc tgatcagtct
     2821 ttttcaacac gtaaaaagcg gaggagtttt gcaattttgt tggttgtaac gatcctccgt
     2881 tgattttggc ctctttctcc atgggcgggc tgggcgtatt tgaagcgtag ggataacagg
     2941 gtaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc
     3001 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     3061 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     3121 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     3181 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     3241 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     3301 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     3361 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     3421 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3481 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     3541 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3601 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     3661 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     3721 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     3781 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     3841 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     3901 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     3961 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     4021 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     4081 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     4141 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     4201 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     4261 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     4321 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     4381 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     4441 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     4501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4561 aaggatcttc gttaactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
     4621 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga
     4681 agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga
     4741 tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc
     4801 cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat
     4861 gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct
     4921 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
     4981 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtccccc
     5041 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
     5101 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
     5161 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta
     5221 tacacagcca gtctgcaggt cgaccatagt gttaac
//
