LOCUS       Exported                5385 bp ds-DNA     circular SYN 16-JUN-2022
DEFINITION  Mammalian expression of rat LC3 fused to mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    pmRFP-LC3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5385)
  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. 2007;3(5):452-60.
  PUBMED    17534139
REFERENCE   2  (bases 1 to 5385)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5385
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        254..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             790..1464
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(920..938)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1188..1207
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1382..1405
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1474..1896
                     /codon_start=1
                     /gene="Map1lc3b"
                     /product="rat LC3 protein"
                     /label=LC3
                     /note="Atg8 family member found on autophagosomes"
                     /translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
                     LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
                     DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
     polyA_signal    2366..2487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2403..2422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2457..2476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2494..2949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2631..2652)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2843..2862
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2976..3080
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3082..3439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3105..3125)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3290..3425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3352..3371
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3474..4268
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3528..3547)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4138..4157
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4456..4475)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4500..4547
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4876..79
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5365..5384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
       61 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      121 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      181 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      781 tccgctagca tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc
      841 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      901 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
      961 tgggacatcc tgtcccctca gttccagtac ggctccaagg cctacgtgaa gcaccccgcc
     1021 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1081 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1141 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1201 aagaagacca tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg
     1261 aagggcgaga tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc
     1321 aagaccacct acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc
     1381 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
     1441 gagggccgcc actccaccgg cgccagatct agaccgtccg agaagacctt caaacagcgc
     1501 cggagcttcg aacaaagagt ggaagatgtc cggctcatcc gggagcagca ccccaccaag
     1561 atcccagtga ttatagagcg atacaagggt gagaagcagc tgcccgtcct ggacaagacc
     1621 aagttccttg tacctgatca cgtgaatatg agcgaactca tcaagataat tagaaggcgc
     1681 ctgcagctca atgctaacca agccttcttc ctcctggtga atgggcacag catggtgagt
     1741 gtgtccacac ccatctctga agtgtacgag agcgagagag atgaagacgg cttcctgtac
     1801 atggtctatg cctcccagga gacgttcggg acagcactgg ctgttacata catgtcagct
     1861 ctgaaggcaa cagcaacagg aagagagcca tgcttgtgac agacatacag ccacttccaa
     1921 ctaaagcaag cctctgctcc ctgctacctg catggagccc actgtgacac tcagaccatc
     1981 cccggtcact cactcactcg tgtctgagaa tctcagtgag agctgcctct gtcacggacg
     2041 gaagccaaca cagccacctc tcgacctggc tccccacagc acccaccctc cctgcatgca
     2101 gctgtccctg ctaaccccca atgttatgtt acactgtgta agtccccact gctgccgtgt
     2161 gtgggttgtg tacgtcgtca tgtccctggt ttataactat ggtgcgaatt ctgcagtcga
     2221 cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca gccataccac
     2281 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2341 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2401 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2461 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     2521 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2581 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2641 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2701 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2761 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2821 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2881 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2941 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3061 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3121 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3181 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3241 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3301 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3361 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3421 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3481 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3541 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3601 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3661 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3721 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3781 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3841 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3901 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3961 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4021 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4081 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4141 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4201 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4261 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4321 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4381 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4441 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4501 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4561 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4621 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4681 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4741 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5341 agagcgcacg agggagcttc cagggggaaa cgcctggtat cttta
//
