LOCUS       40924_30510        4107 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pME-mCherry-T2A-EGFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4107)
  AUTHORS   Hall TE, Currie PD.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-SEP-2011) Australian Regenerative Medicine Institute, 
            Monash University, Building 75, Melbourne, VIC 3800, Australia
REFERENCE   2  (bases 1 to 4107)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4107)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (27-SEP-2011) Australian Regenerative Medicine Institute, Monash 
            University, Building 75, Melbourne, VIC 3800, Australia"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(387..473)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             674..1381
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    1382..1435
                     /label=T2A viral peptide
                     /note="T2A viral peptide"
     CDS             1382..1435
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1436..2152
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2168..2230
                     /label=CAAX membrane localisation signal (from H-ras)
                     /note="CAAX membrane localisation signal (from H-ras)"
     protein_bind    complement(2232..2331)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2349..2367)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2372..2388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2501..3307
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3457..4045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg accatggtga gcaagggcga ggaggacaac atggccatca tcaaggagtt
      721 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      781 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      841 cggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
      901 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
      961 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1021 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1081 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     1141 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     1201 ccactacgac gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     1261 cgcctacaac gtcaacatca agctggacat cacctcccac aacgaggact acaccatcgt
     1321 ggaacagtac gagcgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1381 ggagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaatggt
     1441 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1501 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1561 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1621 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1681 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1741 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1801 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1861 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1921 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1981 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2041 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2101 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggaggagg
     2161 aagatctaag ctgaaccctc ctgatgagag tggccccggc tgcatgagct gcaagtgtgt
     2221 gctctcctga gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2281 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2341 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2401 cgtgtctcaa aatctctgat gttacattgc acaagataaa ataatatcat catgaacaat
     2461 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2521 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2581 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2641 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2701 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2761 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2821 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2881 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2941 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3001 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3061 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3121 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3181 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3241 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3301 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3361 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4081 cttttgctgg ccttttgctc acatgtt
//
