LOCUS       sequence-443186-        7949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of WIPI2b-mRuby3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7949)
  AUTHORS   Shinoda S, Sakai Y, Matsui T, Uematsu M, Koyama-Honda I, Sakamaki 
            JI, Yamamoto H, Mizushima N
  TITLE     Syntaxin 17 recruitment to mature autophagosomes is temporally 
            regulated by PI4P accumulation.
  JOURNAL   Elife. 2024 Jun 4;12:RP92189. doi: 10.7554/eLife.92189.
  PUBMED    38831696
REFERENCE   2  (bases 1 to 7949)
  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: "Elife."; 
            date: "2024-06-4"; volume: "12:RP92189. doi"; pages: " 
            10.7554/eLife.92189"
COMMENT     Sequence Label: pMRXIB-WIPI2b-mRuby3
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1507..1529
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1545..1561
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1899..1915
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    4085..4651
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4101..4653
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4245..4262)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4472..4491
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             5275..5744
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5874..5891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6045..6633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6125..6144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6797..7588,7589..7657))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7420..7439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7658..7762)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7830..7848
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatta
     1981 attgaattca cgcgttcgcg agttaacgga tccatgaacc tggcgagcca gagcggggag
     2041 gccggcgccg gccagctgct cttcgccaac ttcaaccagg acaacacatc cctagctgtt
     2101 ggtagtaagt ccggttataa atttttctcc ctttcttctg tggataagct ggaacagatc
     2161 tatgaatgca ccgatacgga agatgtgtgc attgtagaga gattgttctc cagcagccta
     2221 gtggccatcg tgagccttaa agcaccaagg aagctaaagg tttgccactt taagaaggga
     2281 actgagatct gcaactacag ctactccaac acgattctgg ctgtgaagct caacaggcag
     2341 aggctgatag tatgcctgga ggagtccctg tacatccaca acattcggga catgaaggtg
     2401 ctgcatacga tcagggagac gcctccaaac cctgcaggcc tgtgtgcgct gtcaatcaac
     2461 aacgacaact gctacttggc gtacccaggg agcgcgacca tcggagaggt gcaggtcttc
     2521 gataccatta atttgagagc tgcaaacatg attccggctc acgacagtcc tttagcggca
     2581 ctggcctttg acgcaagtgg aactaaactt gccacggctt cggagaaggg gaccgtgatt
     2641 agggtatttt ccattccaga aggacaaaaa ctctttgagt ttcggagagg agtaaagagg
     2701 tgcgtgagca tctgctccct ggccttcagc atggacggca tgttcctctc cgcctccagc
     2761 aacactgaga ccgtgcacat cttcaaactc gagactgtga aagaaaaacc cccagaggag
     2821 cccaccacct ggaccgggta cttcgggaaa gtgctcatgg cctccaccag ctacctgcct
     2881 tcccaagtga cagaaatgtt caaccagggc agagccttcg ccacggtccg cctgccattc
     2941 tgcggccaca aaaacatctg ctcgctagcc acaattcaga agatcccgcg gttgttggtg
     3001 ggtgccgccg acgggtacct gtacatgtac aacctggacc cccaggaggg cggcgagtgt
     3061 gccctgatga agcagcaccg gctggacggc agtctggaaa cgaccaatga gatcttggac
     3121 tctgcctctc acgactgccc cttagtcact cagacatacg gcgcagctgc aggaaaaggt
     3181 acttacgtgc cttcatcccc aacgagactt gcctacacag acgacctggg tgctgtgggt
     3241 ggcgcctgcc tggaggacga ggccagcgcc ctgcgcctgg atgaggacag cgagcacccg
     3301 cccatgattc ttcggactga cgaattcatg gtgggatccg tgagcaaggg cgaggagctc
     3361 atcaaggaga acatgcggat gaaagtcgtc atggaaggct ctgtcaatgg gcaccagttc
     3421 aagtgtaccg gagaagggga agggcgtccc tatgaaggcg tacaaaccat gcgcatcaaa
     3481 gtgatagagg gcgggccact cccttttgcc ttcgacattc tggctacctc cttcatgtat
     3541 ggcagtcgga cattcatcaa gtatcccgca gacattccgg acttcttcaa acagtccttt
     3601 ccagaaggct ttacatggga gagggtgact aggtacgaag atggcggagt ggttactgtg
     3661 actcaggaca cgtcattgga ggacggagag ctggtgtaca atgtgaaagt acgcggagtg
     3721 aactttccca gcaatggtcc tgttatgcag aagaaaacca agggttggga acccaatacg
     3781 gagatgatgt atccagctga tggtggcctg aggggataca cagacatagc cctgaaagtc
     3841 gatggaggtg gccatctgca ctgcaacttt gtgaccacct acagaagcaa gaaaactgtg
     3901 gggaacatta agatgcctgg agtgcatgca gtcgatcaca gacttgagag aatcgaggag
     3961 tctgataacg agacatacgt ggtacaacga gaagttgccg ttgcgaagta cagcaatctg
     4021 ggcgggggca tggacgagct gtacaagctc gagtaaggat ccctcgagcg gccgctacgt
     4081 aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4141 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4201 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4261 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     4321 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     4381 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     4441 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     4501 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     4561 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     4621 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtc atgaaaacat
     4681 ttaacatttc tcaacaagat ctagaattag tagaagtagc gacagagaag attacaatgc
     4741 tttatgagga taataaacat catgtgggag cggcaattcg tacgaaaaca ggagaaatca
     4801 tttcggcagt acatattgaa gcgtatatag gacgagtaac tgtttgtgca gaagccattg
     4861 cgattggtag tgcagtttcg aatggacaaa aggattttga cacgattgta gctgttagac
     4921 acccttattc tgacgaagta gatagaagta ttcgagtggt aagtccttgt ggtatgtgta
     4981 gggagttgat ttcagactat gcaccagatt gttttgtgtt aatagaaatg aatggcaagt
     5041 tagtcaaaac tacgattgaa gaactcattc cactcaaata tacccgaaat taacgacccc
     5101 atggctcgac gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     5161 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     5221 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     5281 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     5341 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     5401 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     5461 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     5521 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     5581 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     5641 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     5701 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     5761 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     5821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6721 aggatcttca cctagatcct tttgcggccg gccgcaaatc aatctaaagt atatatgagt
     6781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7381 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     7441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7801 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     7861 actcgaggag cttggcccat tgcatacgtt gtatccatat cataatatgt acatttatat
     7921 tggctcatgt ccaacattac cgccatgtt
//
