LOCUS       sequence-313934-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  AUTHORS   Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, 
            Tokuhisa T, Ohsumi Y, Yoshimori T
  TITLE     Dissection of autophagosome formation using Apg5-deficient mouse 
            embryonic stem cells.
  JOURNAL   J Cell Biol. 2001 Feb 19. 152(4):657-68.
  PUBMED    11266458
REFERENCE   2  (bases 1 to 5586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2001 Feb 19. 152(4):657-68."
COMMENT     Sequence Label: pEGFP-C1-mApg5
FEATURES             Location/Qualifiers
     source          1..5586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2151..2167)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2152..2168)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2314..2435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2351..2370)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2405..2424
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2442..2897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2579..2600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2791..2810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2924..3028
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3030..3387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3030..3221
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3053..3073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3238..3373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3422..4216
                     /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             3422..4216
                     /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(3476..3495)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4086..4105
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4404..4423)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4448..4495
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4824..5412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5313..5332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atcccccggg atgacagatg acaaagatgt gcttcgagat
     1321 gtgtggtttg gacgaattcc aacttgcttt actctctatc aggatgagat aactgaaaga
     1381 gaagcagaac catactattt gcttttgcca agagtcagct atttgacgtt ggtaactgac
     1441 aaagtgaaaa agcactttca gaaggttatg agacaagaag atgttagtga gatatggttt
     1501 gaatatgaag gcacacccct gaaatggcat tatccaattg gtttactatt tgatcttctt
     1561 gcatcaagtt cagctcttcc ttggaacatc acagtacatt tcaagagttt tccagaaaag
     1621 gaccttctac actgtccatc caaggatgcg gttgaggctc actttatgtc gtgtatgaaa
     1681 gaagctgatg ctttaaagca taaaagtcaa gtgatcaacg aaatgcagaa aaaagaccac
     1741 aagcagctct ggatgggact gcagaatgac agatttgacc agttttgggc catcaaccgg
     1801 aaactcatgg aatatcctcc agaagaaaat ggatttcgtt atatcccctt tagaatatat
     1861 cagaccacga cggagcggcc tttcatccag aagctgttcc ggcctgtggc cgcagatgga
     1921 cagctgcaca cacttggaga tctcctcaga gaagtctgtc cttccgcagt cgcccctgaa
     1981 gatggagaga agaggagcca ggtgatgatt cacgggatag agccaatgct ggaaacccct
     2041 ctgcagtggc tgagcgagca tctgagctac ccagataact ttcttcatat tagcattgtc
     2101 ccccagccaa cagattgacc cgggctgcag gaattcgata tcaagcttat cgataccgtc
     2161 gacctcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     2221 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2281 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2341 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2401 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     2461 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2521 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2581 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2641 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2701 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2761 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2821 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2881 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2941 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3001 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3301 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3361 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3421 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3481 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3541 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3601 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3661 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3721 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3781 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3841 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3901 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3961 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4021 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4081 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4141 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4201 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4261 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4321 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4381 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4441 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4501 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4561 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4621 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4681 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4741 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4801 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4921 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4981 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5041 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5101 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5161 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5221 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5281 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5341 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5401 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5461 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5521 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5581 gttccg
//
