LOCUS       EGFP-LC3                5113 bp ds-DNA     circular SYN 16-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    EGFP-LC3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5113)
  AUTHORS   Jackson WT, Giddings TH, Taylor MP, Mulinyawe S, Rabinovitch M, 
            Kopito RR, Kirkegaard K
  TITLE     Subversion of cellular autophagosomal machinery by RNA viruses.
  JOURNAL   PLoS Biol. 2005 May . 3(5):e156.
  PUBMED    15884975
REFERENCE   2  (bases 1 to 5113)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 16, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..5113
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(24..381)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      38..173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(92..111)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     338..358
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(383..487)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      514..969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(601..620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     811..832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    976..1097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(987..1006)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1041..1060
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1261..1635)
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVYASQETFGMKLSV"
     CDS             complement(1669..2385)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(1669..2385)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1711..1732)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2060..2079
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2319..2340
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2382..2391
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(2430..2633)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2459..2479)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2634..2937
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3112..3700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3192..3211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4029..4076
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4101..4120
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4308..5102)
                     /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(4419..4438)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5029..5048
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac
       61 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
      121 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
      181 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
      241 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
      301 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
      361 agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc atttatcagg
      421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      481 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
      541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      721 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      841 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
      961 ttacaattta cgcgttaaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1021 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1081 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1141 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     1201 atcagttatc tagatccggt ggatcccggg cccgcggtac cgtcgactgc agaattctta
     1261 cactgacaat ttcatcccga acgtctcctg ggaggcatag accatgtaca ggaatccatc
     1321 ttcatctttc tcactctcat acacctctga gattggtgtg gagacgctga ccatgctgtg
     1381 tccgttcacc aacaggaaga aggcctgatt agcattgagc tgtaagcgcc ttctaattat
     1441 cttgatgagc tcactcatgt tgacatggtc aggtacaagg aactttgttt tatccagaac
     1501 aggaagctgc ttctcaccct tgtatcgttc tattatcacc gggattttgg ttggatgctg
     1561 ctctcgaata agtcggacat cttctactct ttgttcgaag gtgcggcgct gcttgaaggt
     1621 cttctccgac ggcatgggaa ttcgaagctt gagctcgaga tctgagtact tgtacagctc
     1681 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     1741 cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag
     1801 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     1861 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     1921 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     1981 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     2041 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     2101 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     2161 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     2221 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     2281 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     2341 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag
     2401 cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca
     2461 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     2521 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     2581 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     2641 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     2701 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     2761 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     2821 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     2881 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     2941 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactaatg
     3001 catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3061 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3121 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3181 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3241 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3301 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3361 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3421 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3481 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3541 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3601 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3841 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     3901 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     3961 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     4021 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     4081 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     4141 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     4201 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     4261 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     4321 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     4381 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     4441 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     4501 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     4561 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     4621 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     4681 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     4741 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     4801 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     4861 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     4921 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     4981 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     5041 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     5101 atgcgaaacg atc
//