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"
     primer_bind     complement(30..49)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(321..425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      452..907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(539..558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     749..770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    914..1035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(925..944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     979..998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1505..1927)
                     /codon_start=1
                     /gene="Map1lc3b"
                     /product="rat LC3 protein"
                     /label=LC3
                     /note="Atg8 family member found on autophagosomes"
                     /translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
                     LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
                     DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
     CDS             complement(1937..2611)
                     /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(1996..2019)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2194..2213)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2463..2481
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(2640..2843)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2669..2689)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2844..3147
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3322..3910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3402..3421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4239..4286
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4311..4330
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4518..5312)
                     /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(4629..4648)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5239..5258
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5347..319)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5361..111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
      361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      421 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
      481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      661 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      781 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      901 acaatttacg cgttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      961 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1021 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1081 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     1141 cagttatcta gatccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcgcacc
     1201 atagttataa accagggaca tgacgacgta cacaacccac acacggcagc agtggggact
     1261 tacacagtgt aacataacat tgggggttag cagggacagc tgcatgcagg gagggtgggt
     1321 gctgtgggga gccaggtcga gaggtggctg tgttggcttc cgtccgtgac agaggcagct
     1381 ctcactgaga ttctcagaca cgagtgagtg agtgaccggg gatggtctga gtgtcacagt
     1441 gggctccatg caggtagcag ggagcagagg cttgctttag ttggaagtgg ctgtatgtct
     1501 gtcacaagca tggctctctt cctgttgctg ttgccttcag agctgacatg tatgtaacag
     1561 ccagtgctgt cccgaacgtc tcctgggagg catagaccat gtacaggaag ccgtcttcat
     1621 ctctctcgct ctcgtacact tcagagatgg gtgtggacac actcaccatg ctgtgcccat
     1681 tcaccaggag gaagaaggct tggttagcat tgagctgcag gcgccttcta attatcttga
     1741 tgagttcgct catattcacg tgatcaggta caaggaactt ggtcttgtcc aggacgggca
     1801 gctgcttctc acccttgtat cgctctataa tcactgggat cttggtgggg tgctgctccc
     1861 ggatgagccg gacatcttcc actctttgtt cgaagctccg gcgctgtttg aaggtcttct
     1921 cggacggtct agatctggcg ccggtggagt ggcggccctc ggcgcgctcg tactgttcca
     1981 cgatggtgta gtcctcgttg tgggaggtga tgtccagctt gatgtcggtc ttgtaggcgc
     2041 cgggcagctg cacgggcttc ttggccatgt aggtggtctt gacctcggcg tcgtagtggc
     2101 cgccgtcctt cagcttcagc ctcatcttga tctcgccctt cagggcgccg tcctcggggt
     2161 acatccgctc ggtggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg
     2221 aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg
     2281 agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga
     2341 agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg
     2401 ccttggagcc gtactggaac tgaggggaca ggatgtccca ggcgaagggc agggggccgc
     2461 ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc
     2521 cctcgatctc gaactcgtgg ccgttcacgg agccctccat gcgcaccttg aagcgcatga
     2581 actccttgat gacgtcctcg gaggaggcca tgctagcgga tctgacggtt cactaaacca
     2641 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     2701 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     2761 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     2821 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     2881 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     2941 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     3001 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     3061 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     3121 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     3181 cccgtaattg attactatta ataactaatg catggcggta atacggttat ccacagaatc
     3241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3841 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3901 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     4081 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     4141 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     4201 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     4261 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     4321 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     4381 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     4441 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     4501 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     4561 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     4621 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4681 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     4741 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     4801 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     4861 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     4921 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     4981 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5041 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5101 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     5161 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5221 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5281 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     5341 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cacta
//
