LOCUS       sequence-453951-        7152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SNAP-Rab7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  AUTHORS   Kurikawa Y, Koyama-Honda I, Tamura N, Koike S, Mizushima N
  TITLE     Organelle landscape analysis using a multiparametric particle-based 
            method.
  JOURNAL   PLoS Biol. 2024 Sep 17;22(9):e3002777. doi: 
            10.1371/journal.pbio.3002777. eCollection 2024 Sep.
  PUBMED    39288101
REFERENCE   2  (bases 1 to 7152)
  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: "PLoS 
            Biol."; date: "2024-09-17"; volume: "22(9):e3002777. doi"; pages: " 
            10.1371/journal.pbio.3002777. eCollection 2024 Sep"
COMMENT     Sequence Label: pMRX-IBU-SNAP-Rab7
FEATURES             Location/Qualifiers
     source          1..7152
                     /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"
     CDS             1994..2539
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    3234..3800
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3250..3802
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3394..3411)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3621..3640
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4478..4947
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5077..5094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5248..5836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5328..5347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6000..6791,6792..6860))
                     /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     6623..6642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6861..6965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7033..7051
                     /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 accatggaca aagactgcga aatgaagcgc accaccctgg atagccctct
     2041 gggcaagctg gaactgtctg ggtgcgaaca gggcctgcac cgtatcatct tcctgggcaa
     2101 aggaacatct gccgccgacg ccgtggaagt gcctgcccca gccgccgtgc tgggcggacc
     2161 agagccactg atgcaggcca ccgcctggct caacgcctac tttcaccagc ctgaggccat
     2221 cgaggagttc cctgtgccag ccctgcacca cccagtgttc cagcaggaga gctttacccg
     2281 ccaggtgctg tggaaactgc tgaaagtggt gaagttcgga gaggtcatca gctacagcca
     2341 cctggccgcc ctggccggca atcccgccgc caccgccgcc gtgaaaaccg ccctgagcgg
     2401 aaatcccgtg cccattctga tcccctgcca ccgggtggtg cagggcgacc tggacgtggg
     2461 gggctacgag ggcgggctcg ccgtgaaaga gtggctgctg gcccacgagg gccacagact
     2521 gggcaagcct gggctgggtg gttcaggctc gggatccggg aattcgggtc tcgagaaaag
     2581 cggccgccac atgacctcta ggaagaaagt gttgctgaag gtcatcatcc tgggggactc
     2641 tggtgttgga aagacctctc tcatgaacca gtatgtgaac aagaagttca gtaaccagta
     2701 caaagccaca ataggagcgg actttctgac caaggaggtg atggtggacg acagacttgt
     2761 taccatgcag atctgggaca cagccggtca agaacggttc cagtctcttg gtgtggcctt
     2821 ctacagaggt gcagattgct gtgttctggt gtttgatgtg actgccccca acactttcaa
     2881 aaccctcgac agctggagag acgagtttct catccaggcc agcccccggg atcccgagaa
     2941 cttccctttt gttgtgttgg gaaacaagat tgacctggaa aacagacaag tggccacaaa
     3001 gagggcacag gcttggtgct acagcaaaaa caacattcct tacttcgaga ccagtgccaa
     3061 ggaggccatc aatgtggagc aggccttcca gacaattgct cggaatgccc ttaaacagga
     3121 aacagaagtg gaactgtaca atgaattccc tgaacccatc aaactggaca agaatgaccg
     3181 ggccaaggcc tccgcagaaa gctgcagttg ttgagagcgg ccgctacgta aattccgccc
     3241 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     3301 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     3361 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     3421 tgaatgtcgt gaaggaagca gttcctctgg aagtttcttg aagacaaaca acgtctgtag
     3481 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     3541 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     3601 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     3661 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     3721 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     3781 tttgaaaaac acgatgataa tatggccaca accgtcgaca tggtcatgaa aacatttaac
     3841 atttctcaac aagatctaga attagtagaa gtagcgacag agaagattac aatgctttat
     3901 gaggataata aacatcatgt gggagcggca attcgtacga aaacaggaga aatcatttcg
     3961 gcagtacata ttgaagcgta tataggacga gtaactgttt gtgcagaagc cattgcgatt
     4021 ggtagtgcag tttcgaatgg acaaaaggat tttgacacga ttgtagctgt tagacaccct
     4081 tattctgacg aagtagatag aagtattcga gtggtaagtc cttgtggtat gtgtagggag
     4141 ttgatttcag actatgcacc agattgtttt gtgttaatag aaatgaatgg caagttagtc
     4201 aaaactacga ttgaagaact cattccactc aaatataccc gaaattaagt cgaccgcccg
     4261 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc gacgataaaa
     4321 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     4381 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     4441 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     4501 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     4561 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4621 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     4681 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4741 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4801 cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     4861 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4921 tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt
     4981 tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg
     5041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5941 ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6001 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6061 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6121 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6181 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6241 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6301 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6361 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6421 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6481 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6541 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6601 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6661 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6721 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6781 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6841 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6961 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7021 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcactatag gagcttggcc
     7081 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat
     7141 taccgccatg tt
//
