LOCUS       sequence-443160-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of mRuby3-STX17TM in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  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 6905)
  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-STX17TM
FEATURES             Location/Qualifiers
     source          1..6905
                     /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    2987..3553
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3003..3555
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3147..3164)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3374..3393
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4231..4700
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4830..4847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5001..5589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5081..5100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5753..6544,6545..6613))
                     /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     6376..6395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6614..6718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6786..6804
                     /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 ccgggaattc gggtctggca gctctgcctg tggcaggtgc
     2761 actcatcggg ggaatggtag ggggtcctat tggcctcctt gcaggcttca aagtggcagg
     2821 aattgcagct gcacttggtg gtggggtgtt gggcttcaca ggtggaaaat tgatacaaag
     2881 aaagaaacag aaaatgatgg agaagctcac ttccagctgt ccagatcttc ccagccaaac
     2941 tgacaagaaa tgcagttgag gtctcgagag cggccgctac gtaaattccg cccccccccc
     3001 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     3061 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     3121 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     3181 cgtgaaggaa gcagttcctc tggaagtttc ttgaagacaa acaacgtctg tagcgaccct
     3241 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     3301 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     3361 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     3421 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     3481 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3541 aacacgatga taatatggcc acaaccgtcg acatggtcat gaaaacattt aacatttctc
     3601 aacaagatct agaattagta gaagtagcga cagagaagat tacaatgctt tatgaggata
     3661 ataaacatca tgtgggagcg gcaattcgta cgaaaacagg agaaatcatt tcggcagtac
     3721 atattgaagc gtatatagga cgagtaactg tttgtgcaga agccattgcg attggtagtg
     3781 cagtttcgaa tggacaaaag gattttgaca cgattgtagc tgttagacac ccttattctg
     3841 acgaagtaga tagaagtatt cgagtggtaa gtccttgtgg tatgtgtagg gagttgattt
     3901 cagactatgc accagattgt tttgtgttaa tagaaatgaa tggcaagtta gtcaaaacta
     3961 cgattgaaga actcattcca ctcaaatata cccgaaatta agtcgaccgc ccgccccacg
     4021 acccgcagcg cccgaccgaa aggagcgcac gaccccatgg ctcgacgata aaataaaaga
     4081 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     4141 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     4201 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     4261 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     4321 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     4381 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     4441 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     4501 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
     4561 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     4621 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     4681 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     4741 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     4801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5401 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     5701 cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5821 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5881 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5941 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6001 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6061 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6361 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6721 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6781 ataaaaatag gcgtatcacg aggccctttc gtcttcacta taggagcttg gcccattgca
     6841 tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc
     6901 atgtt
//
