LOCUS       ptfLC3.        6136 bp DNA     circular SYN 10-JAN-2024
DEFINITION  Mammalian expression of rat LC3 fused to mRFP and EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    ptfLC3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6136)
  AUTHORS   Kimura S, Noda T, Yoshimori T
  TITLE     Dissection of the autophagosome maturation process by a novel 
            reporter protein, tandem fluorescent-tagged LC3.
  JOURNAL   Autophagy.   . 3(5):452-60.
  PUBMED    17534139
REFERENCE   2  (bases 1 to 6136)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6136)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Autophagy. 
             . 3(5):452-60."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        108..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             651..1325
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     CDS             1344..2060
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             2079..2501
                     /codon_start=1
                     /gene="Map1lc3b"
                     /product="rat LC3 protein"
                     /label=LC3
                     /note="Atg8 family member found on autophagosomes"
                     /translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
                     LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
                     DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
     polyA_signal    2971..3092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3099..3554)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3581..3685
                     /label=AmpR promoter
     promoter        3687..4044
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4079..4870
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     primer_bind     complement(5061..5080)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5105..5152
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5481..6069
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctggttt agtgaaccgt cagatccgct agcggccacc atggccttct
      661 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
      721 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
      781 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
      841 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
      901 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
      961 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1021 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1081 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
     1141 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
     1201 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
     1261 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg
     1321 gcgccggtgg accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1381 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1441 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1501 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1561 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1621 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1681 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1741 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1801 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1861 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1921 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1981 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2041 gcatggacga gctgtacaag tccggactca gatctagacc gtccgagaag accttcaaac
     2101 agcgccggag cttcgaacaa agagtggaag atgtccggct catccgggag cagcacccca
     2161 ccaagatccc agtgattata gagcgataca agggtgagaa gcagctgccc gtcctggaca
     2221 agaccaagtt ccttgtacct gatcacgtga atatgagcga actcatcaag ataattagaa
     2281 ggcgcctgca gctcaatgct aaccaagcct tcttcctcct ggtgaatggg cacagcatgg
     2341 tgagtgtgtc cacacccatc tctgaagtgt acgagagcga gagagatgaa gacggcttcc
     2401 tgtacatggt ctatgcctcc caggagacgt tcgggacagc actggctgtt acatacatgt
     2461 cagctctgaa ggcaacagca acaggaagag agccatgctt gtgacagaca tacagccact
     2521 tccaactaaa gcaagcctct gctccctgct acctgcatgg agcccactgt gacactcaga
     2581 ccatccccgg tcactcactc actcgtgtct gagaatctca gtgagagctg cctctgtcac
     2641 ggacggaagc caacacagcc acctctcgac ctggctcccc acagcaccca ccctccctgc
     2701 atgcagctgt ccctgctaac ccccaatgtt atgttacact gtgtaagtcc ccactgctgc
     2761 cgtgtgtggg ttgtgtacgt cgtcatgtcc ctggtttata actatggtgc gaattctgca
     2821 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     2881 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2941 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3061 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3121 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3181 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3241 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3301 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3361 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3421 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3481 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3541 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3661 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3721 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4081 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4141 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4201 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4261 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4321 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4381 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4441 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4501 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4561 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4621 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4681 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4741 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4801 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4861 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4921 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4981 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5041 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5101 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5161 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5221 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5281 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5341 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5401 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5461 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5521 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5581 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5641 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5701 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5761 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5821 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5881 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5941 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6001 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6061 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6121 gctcacatgt tctttc
//
