LOCUS       sequence-166345-        5867 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 5867)
  AUTHORS   Fujita N, Hayashi-Nishino M, Fukumoto H, Omori H, Yamamoto A, Noda 
            T, Yoshimori T
  TITLE     An Atg4B mutant hampers the lipidation of LC3 paralogues and causes 
            defects in autophagosome closure.
  JOURNAL   Mol Biol Cell. 2008 Nov . 19(11):4651-9.
  PUBMED    18768752
REFERENCE   2  (bases 1 to 5867)
  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: "Mol Biol 
            Cell. 2008 Nov . 19(11):4651-9."
COMMENT     Sequence Label: pmStrawberry-Atg4BC74A
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             552..1259
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mStrawberry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(697..715)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     965..984
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2610..2731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2647..2666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2701..2720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2738..3193)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2875..2896)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3087..3106
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3220..3324
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3326..3683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3326..3517
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3349..3369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3534..3669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3718..4512
                     /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             3718..4512
                     /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(3772..3791)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4382..4401
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4700..4719)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4744..4791
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5120..5708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5609..5628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag catggtgagc aagggcgagg agaataacat ggccatcatc aaggagttca
      601 tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      661 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      721 gccccctgcc cttcgcctgg gacatcctaa cccccaactt cacctacggc tccaaggcct
      781 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      841 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      901 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
      961 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1021 ccgaggacgg cgccctgaag ggcgagatca agatgaggct gaagctgaag gacggcggcc
     1081 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1141 cctacatcgt cggcatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1201 aactgtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga
     1261 gatccatgga cgcagccact ttgacatatg atactctccg gtttgctgaa tttgaagatt
     1321 tccctgagac ctcagagcct gtttggattc tgggcagaaa atacagcatt ttcacagaga
     1381 aggacgaaat cttgtctgat gttgcatcca gactttggtt tacatacagg agaaactttc
     1441 cagctattgg gggaactggc cctacttcag acacaggctg gggtgcaatg cttcggtgtg
     1501 gacagatgat ctttgcccag gccctggtat gccggcactt aggtcgagat tggaggtgga
     1561 ctcagcggaa gaggcagcct gacagctact ttaatgtcct caatgctttc ctcgacagga
     1621 aggacagcta ctattccatc catcagatag cgcaaatggg agttggcgaa ggcaagtcta
     1681 taggccagtg gtacgggccg aacactgttg cccaggtcct caagaaactt gctgtattcg
     1741 acacatggag ttccttggct gttcacatag caatggacaa cactgtggtg atggaggaaa
     1801 tcagaaggtt atgcagggcc aaccttccct gtgttggggc ggctgcactt cctactgatt
     1861 cagagaggca ctgtaatggg ttccctgctg gagctgaagt caccaacagg ccatcggctt
     1921 ggagaccact agtgcttctc atccctctcc gcctgggact gacagacatc aatgaggcct
     1981 atgtggagac gctgaagcac tgtttcatga tgccccagtc cctgggtgtt attggaggga
     2041 agcccaacag tgcccactac tttattggct atgttgggga ggagctcatc tatctggacc
     2101 cccacactac acagcctgca gtggagctga ctgacagctg cttcatcccc gacgagagct
     2161 tccactgcca gcaccctccc tctaggatgg gcattggaga gcttgacccg tccattgctg
     2221 tggggttttt ctgtaagaag gaggaagact ttaacgactg gtgtcagcaa gtcaaaaagc
     2281 tatcccagct tggaggtgcc ctgcccatgt ttgagctggt ggaacagcag ccttcccatc
     2341 tggcctgcca ggatgtcctg aacctgtccc tagactcttc tgatgtagag agactggaga
     2401 ggttttttga ctctgaggat gaagactttg aaatcttatc cctctgaggt accgatatcc
     2461 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     2521 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2581 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2641 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2701 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     2761 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2821 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2881 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2941 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3001 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3061 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3121 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3181 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3241 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3301 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3361 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3421 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3541 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3601 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3661 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3721 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3781 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3841 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     3901 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3961 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4021 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4081 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4141 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4201 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4261 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4321 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4381 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4441 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4501 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4561 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4621 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4681 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4741 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4801 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4861 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     4921 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     4981 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5041 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5761 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     5821 attagttatt aatagtaatc aattacgggg tcattagttc atagccc
//
