LOCUS       sequence-453950-        7491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Venus-GS27 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7491)
  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 7491)
  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-IPU-Venus-GS27
FEATURES             Location/Qualifiers
     source          1..7491
                     /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      1989..1998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1995..1997,1998..2000,2001..2711)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2040..2061)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2301..2320)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2648..2669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3402..3968
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3418..3970
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3562..3579)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3789..3808
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3988..4587
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3988..4007)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4484..4504
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4817..5286
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5416..5433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5587..6175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5667..5686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6339..7130,7131..7199))
                     /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     6962..6981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7200..7304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7372..7390
                     /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 aaagcttggc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2041 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2101 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     2161 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     2221 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2281 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2341 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2401 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     2461 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     2521 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2581 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     2641 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2701 agctgtacaa gggttcaggc tcgggatccg ggaattcgat ggatcccctg ttccagcaaa
     2761 cgcacaagca ggtccacgag atccagtctt gcatgggacg cctggagacg gcagacaagc
     2821 agtctgtgca catagtagaa aacgaaatcc aagcaagcat agaccagata ttcagccgtc
     2881 tagaacgtct ggagattttg tccagcaagg agccccctaa caaaaggcaa aatgccaaac
     2941 ttcgggttga ccagttaaag tatgatgtcc agcacctgca gactgcgctc agaaacttcc
     3001 agcatcggcg ccatgcaagg gagcagcagg agagacagcg agaagagctt ctgtctcgaa
     3061 ccttcaccac taacgactct gacaccacca taccaatgga cgaatcactg cagtttaact
     3121 cctccctcca gaaagttcac aacggcatgg atgacctcat tttagatggg cacaatattt
     3181 tagatggact gaggacccag agactgacct tgaaggggac tcagaagaag atccttgaca
     3241 ttgccaacat gctgggcttg tccaacacag tgatgcggct catcgagaag cgggctttcc
     3301 aggacaagta ctttatgata ggtgggatgc tgctgacctg tgtggtcatg ttcctcgtgg
     3361 tgcagtacct gacatgactc gagagcggcc gctacgtaaa ttccgccccc ccccctaacg
     3421 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3481 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3541 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3601 aggaagcagt tcctctggaa gtttcttgaa gacaaacaac gtctgtagcg accctttgca
     3661 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3721 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3781 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3841 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3901 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3961 gatgataata tggccacaac cgtcgacatg accgagtaca agcccacggt gcgcctcgcc
     4021 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     4081 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4141 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4201 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4261 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     4321 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4381 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     4441 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4501 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4561 tgcatgaccc gcaagcccgg tgcctgagtc gaccgcccgc cccacgaccc gcagcgcccg
     4621 accgaaagga gcgcacgacc ccatggctcg acgataaaat aaaagatttt atttagtctc
     4681 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     4741 ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca
     4801 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     4861 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     4921 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     4981 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     5041 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     5101 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgatt
     5161 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     5221 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     5281 ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa
     5341 atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6001 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6241 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa
     6301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     6961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7321 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     7381 atcacgaggc cctttcgtct tcactatagg agcttggccc attgcatacg ttgtatccat
     7441 atcataatat gtacatttat attggctcat gtccaacatt accgccatgt t
//
