LOCUS       sequence-403660-        4685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ash-tagged GABARAP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4685)
  AUTHORS   Ishimura R, El-Gowily AH, Noshiro D, Komatsu-Hirota S, Ono Y, Shindo
            M, Hatta T, Abe M, Uemura T, Lee-Okada HC, Mohamed TM, Yokomizo T, 
            Ueno T, Sakimura K, Natsume T, Sorimachi H, Inada T, Waguri S, Noda 
            NN, Komatsu M
  TITLE     The UFM1 system regulates ER-phagy through the ufmylation of CYB5R3.
  JOURNAL   Nat Commun. 2022 Dec 21;13(1):7857. doi: 10.1038/s41467-022-35501-0.
  PUBMED    36543799
REFERENCE   2  (bases 1 to 4685)
  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: "Nat 
            Commun."; date: "2022-12-21"; volume: "13(1):7857. doi"; pages: " 
            10.1038/s41467-022-35501-0"
COMMENT     Sequence Label: pAsh-MCL-GABARAP
FEATURES             Location/Qualifiers
     source          1..4685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    869..990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(906..925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     960..979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(997..1452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1134..1155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1346..1365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1479..1583
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1585..1942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1585..1776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1608..1628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1976..2770
                     /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             1976..2770
                     /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(2030..2049)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2640..2659
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2958..2977)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3002..3049
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3378..3966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3867..3886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4141..4444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4445..4648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4599..4619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4680..4685,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggcgtcgc tcaccgtgaa ggcctacctt ctgggcaagg aggacgcggc gcgcgagatt
       61 cgccgcttca gcttctgttg cagccccgag cctgaggcgg aagccgaggc tgcggcgggt
      121 ccgggaccct gcgagcggct gctgagccgg gtggccgccc tgttccccgc gctgcggcct
      181 ggcggcttcc aggcgcacta ccgcgatgag gacggggact tggttgcctt ttccagtgac
      241 gaggaattga caatggccat gtcctacgtg aaggatgaca tcttccgaat ctacattaaa
      301 gagaaaaccg gtaattccgc tgacggcggc ggaggatcgg gtggtagtgg tggttcagga
      361 ggaggatcga cccaaggagg atccatgaag ttcgtgtaca aagaagagca tccgttcgag
      421 aagcgccgct ctgagggcga gaagatccga aagaaatacc cggaccgggt gccggtgata
      481 gtagaaaagg ctcccaaagc tcggatagga gacctggaca aaaagaaata cctggtgcct
      541 tctgatctca cagttggtca gttctacttc ttgatccgga agcgaattca tctccgagct
      601 gaggatgcct tgtttttctt tgtcaacaat gtcattccac ccaccagtgc cacaatgggt
      661 cagctgtacc aggaacacca tgaagaagac ttctttctct acattgccta cagtgacgaa
      721 agtgtctacg gttgaaagct tgaataagcg gccgcgactc tagatcataa tcagccatac
      781 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
      841 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
      901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
      961 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1021 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1081 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1141 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1201 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1261 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1321 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1381 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1441 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1501 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1561 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     1621 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1681 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1741 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     1801 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     1861 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     1921 tggaggccta ggcttttgca aagatcatca agagacagga tgaggatcgt ttcgcatgat
     1981 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2041 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2101 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     2161 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2221 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2281 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2341 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2401 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2461 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     2521 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2581 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2641 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2701 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     2761 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     2821 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     2881 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     2941 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3001 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     3061 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     3121 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     3181 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     3241 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3301 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3361 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3421 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3481 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3541 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3601 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3661 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3721 agtttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3781 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3841 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3901 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3961 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4021 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     4081 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     4141 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     4201 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     4261 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     4321 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     4381 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     4441 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     4501 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     4561 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     4621 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcattg
     4681 ccacc
//
