LOCUS       sequence-443436-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of mRuby3-CERT(PHD) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  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 7044)
  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-mRuby3-CERT(PHD)
FEATURES             Location/Qualifiers
     source          1..7044
                     /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"
     regulatory      1988..1997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3126..3692
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3142..3694
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3286..3303)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3513..3532
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4370..4839
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4969..4986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5140..5728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5220..5239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5892..6683,6684..6752))
                     /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     6515..6534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6753..6857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6925..6943
                     /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 ccatcgatat
     1981 aaagcttgcc accatggtga gcaagggcga ggagctcatc aaggagaaca tgcggatgaa
     2041 agtcgtcatg gaaggctctg tcaatgggca ccagttcaag tgtaccggag aaggggaagg
     2101 gcgtccctat gaaggcgtac aaaccatgcg catcaaagtg atagagggcg ggccactccc
     2161 ttttgccttc gacattctgg ctacctcctt catgtatggc agtcggacat tcatcaagta
     2221 tcccgcagac attccggact tcttcaaaca gtcctttcca gaaggcttta catgggagag
     2281 ggtgactagg tacgaagatg gcggagtggt tactgtgact caggacacgt cattggagga
     2341 cggagagctg gtgtacaatg tgaaagtacg cggagtgaac tttcccagca atggtcctgt
     2401 tatgcagaag aaaaccaagg gttgggaacc caatacggag atgatgtatc cagctgatgg
     2461 tggcctgagg ggatacacag acatagccct gaaagtcgat ggaggtggcc atctgcactg
     2521 caactttgtg accacctaca gaagcaagaa aactgtgggg aacattaaga tgcctggagt
     2581 gcatgcagtc gatcacagac ttgagagaat cgaggagtct gataacgaga catacgtggt
     2641 acaacgagaa gttgccgttg cgaagtacag caatctgggc gggggcatgg acgagctgta
     2701 caagggttca ggctcgggat ccatgtcgga taatcagagc tggaactcgt cgggctcgga
     2761 ggaggatcca gagacggagt ctgggccgcc tgtggagcgc tgcggggtcc tcagtaagtg
     2821 gacaaactac attcatgggt ggcaggatcg ttgggtagtt ttgaaaaata atgctctgag
     2881 ttactacaaa tctgaagatg aaacagagta tggctgcaga ggatccatct gtcttagcaa
     2941 ggctgtcatc acacctcacg attttgatga atgtcgattt gatattagtg taaatgatag
     3001 tgtttggtat cttcgtgctc aggatccaga tcatagacag caatggatag atgccattga
     3061 acagcacaag actgaatctg gatatggatg agaattcggg tctcgagagc ggccgctacg
     3121 taaattccgc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     3181 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     3241 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     3301 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagtttct tgaagacaaa
     3361 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3421 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     3481 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3541 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     3601 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     3661 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccgtcga catggtcatg
     3721 aaaacattta acatttctca acaagatcta gaattagtag aagtagcgac agagaagatt
     3781 acaatgcttt atgaggataa taaacatcat gtgggagcgg caattcgtac gaaaacagga
     3841 gaaatcattt cggcagtaca tattgaagcg tatataggac gagtaactgt ttgtgcagaa
     3901 gccattgcga ttggtagtgc agtttcgaat ggacaaaagg attttgacac gattgtagct
     3961 gttagacacc cttattctga cgaagtagat agaagtattc gagtggtaag tccttgtggt
     4021 atgtgtaggg agttgatttc agactatgca ccagattgtt ttgtgttaat agaaatgaat
     4081 ggcaagttag tcaaaactac gattgaagaa ctcattccac tcaaatatac ccgaaattaa
     4141 gtcgaccgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc
     4201 tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     4261 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     4321 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
     4381 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
     4441 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4501 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
     4561 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     4621 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     4681 caacccctca ctcggcgcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     4741 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     4801 cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt
     4861 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     4921 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4981 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5581 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5821 cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact
     5881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6481 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     6541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6901 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcactat
     6961 aggagcttgg cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
     7021 catgtccaac attaccgcca tgtt
//
