LOCUS       sequence-201220-        5113 bp    DNA     circular SYN 03-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Oct 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Biol. 
            2005 May . 3(5):e156."
COMMENT     Sequence Label: EGFP-LC3
FEATURES             Location/Qualifiers
     source          1..5113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..250
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     207..227
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(252..356)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      383..838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(470..489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     680..701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    845..966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(856..875)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     910..929
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1130..1504)
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVYASQETFGMKLSV"
     CDS             complement(join(1538..2248,2249..2251,2252..2254))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1538..2248,2249..2251,2252..2254))
                     /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(1580..1601)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1929..1948
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2188..2209
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2251..2260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2299..2502)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2328..2348)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2503..2806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2981..3569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3061..3080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3898..3945
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3970..3989
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4177..4971)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4177..4971)
                     /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(4288..4307)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4898..4917
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5006..5113,1..250))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5020..5113,1..42)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
       61 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      121 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
      181 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
      241 tccgcctcag gactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      301 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      361 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      421 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      481 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      541 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      601 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      661 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      721 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      781 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttac
      841 gcgttaagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
      901 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
      961 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1021 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagttatct
     1081 agatccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcttac actgacaatt
     1141 tcatcccgaa cgtctcctgg gaggcataga ccatgtacag gaatccatct tcatctttct
     1201 cactctcata cacctctgag attggtgtgg agacgctgac catgctgtgt ccgttcacca
     1261 acaggaagaa ggcctgatta gcattgagct gtaagcgcct tctaattatc ttgatgagct
     1321 cactcatgtt gacatggtca ggtacaagga actttgtttt atccagaaca ggaagctgct
     1381 tctcaccctt gtatcgttct attatcaccg ggattttggt tggatgctgc tctcgaataa
     1441 gtcggacatc ttctactctt tgttcgaagg tgcggcgctg cttgaaggtc ttctccgacg
     1501 gcatgggaat tcgaagcttg agctcgagat ctgagtactt gtacagctcg tccatgccga
     1561 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     1621 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
     1681 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     1741 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat
     1801 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     1861 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     1921 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     1981 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     2041 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     2101 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     2161 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     2221 ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtagc gctagcggat
     2281 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     2341 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     2401 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     2461 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     2521 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     2581 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     2641 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     2701 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     2761 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     2821 tgggctatga actaatgacc ccgtaattga ttactattaa taactaatgc atggcggtaa
     2881 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2941 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3001 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3061 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3121 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3181 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3241 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3301 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3361 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3421 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3481 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3601 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3661 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3721 agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt
     3781 cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat
     3841 ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa
     3901 caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc
     3961 cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact
     4021 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     4081 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     4141 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4201 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     4261 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     4321 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     4381 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     4441 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     4501 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     4561 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     4621 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     4681 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     4741 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     4801 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4861 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4921 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     4981 tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc
     5041 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     5101 ttagtcagcc atg
//
