LOCUS       sequence-429123-        8354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human MPL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8354)
  TITLE     Masubuchi et al., 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8354)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMSCV-IRES-DsRed/MPL
FEATURES             Location/Qualifiers
     source          1..8354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    169..510
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             577..993
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     923..945
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     961..977
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2946..3495
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3090..3107)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3317..3336
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3498..4175
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(3628..3646)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3678..3698)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3896..3915
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4090..4113
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     LTR             4337..4851
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5013..5029
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5037..5043,5044..5061,5062..5067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5082..5103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5220..5237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5391..5979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5471..5490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6150..6941,6942..7010))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6773..6792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7011..7115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7183..7201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7239..7261)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7361..7380
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7574..7596
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7724..7743
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7943..8354,1..105)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
       61 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
      121 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
      181 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
      241 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      301 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
      361 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
      421 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
      481 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
      541 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
      601 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
      661 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
      721 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
      781 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
      841 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
      901 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
      961 ctttatccag ccctcactcc ttctctaggc gccggaattc gccaccatgc cctcctgggc
     1021 cctcttcatg gtcacctcct gcctcctcct ggcccctcaa aacctggccc aagtcagcag
     1081 ccaagatgtc tccttgctgg catcagactc agagcccctg aagtgtttct cccgaacatt
     1141 tgaggacctc acttgcttct gggatgagga agaggcagcg cccagtggga cataccagct
     1201 gctgtatgcc tacccgcggg agaagccccg tgcttgcccc ctgagttccc agagcatgcc
     1261 ccactttgga acccgatacg tgtgccagtt tccagaccag gaggaagtgc gtctcttctt
     1321 tccgctgcac ctctgggtga agaatgtgtt cctaaaccag actcggactc agcgagtcct
     1381 ctttgtggac agtgtaggcc tgccggctcc ccccagtatc atcaaggcca tgggtgggag
     1441 ccagccaggg gaacttcaga tcagctggga ggagccagct ccagaaatca gtgatttcct
     1501 gaggtacgaa ctccgctatg gccccagaga tcccaagaac tccactggtc ccacggtcat
     1561 acagctgatt gccacagaaa cctgctgccc tgctctgcag aggcctcact cagcctctgc
     1621 tctggaccag tctccatgtg ctcagcccac aatgccctgg caagatggac caaagcagac
     1681 ctccccaagt agagaagctt cagctctgac agcagagggt ggaagctgcc tcatctcagg
     1741 actccagcct ggcaactcct actggctgca gctgcgcagc gaacctgatg ggatctccct
     1801 cggtggctcc tggggatcct ggtccctccc tgtgactgtg gacctgcctg gagatgcagt
     1861 ggcacttgga ctgcaatgct ttaccttgga cctgaagaat gttacctgtc aatggcagca
     1921 acaggaccat gctagctccc aaggcttctt ctaccacagc agggcacggt gctgccccag
     1981 agacaggtac cccatctggg agaactgcga agaggaagag aaaacaaatc caggactaca
     2041 gaccccacag ttctctcgct gccacttcaa gtcacgaaat gacagcatta ttcacatcct
     2101 tgtggaggtg accacagccc cgggtactgt tcacagctac ctgggctccc ctttctggat
     2161 ccaccaggct gtgcgcctcc ccaccccaaa cttgcactgg agggagatct ccagtgggca
     2221 tctggaattg gagtggcagc acccatcgtc ctgggcagcc caagagacct gttatcaact
     2281 ccgatacaca ggagaaggcc atcaggactg gaaggtgctg gagccgcctc tcggggcccg
     2341 aggagggacc ctggagctgc gcccgcgatc tcgctaccgt ttacagctgc gcgccaggct
     2401 caacggcccc acctaccaag gtccctggag ctcgtggtcg gacccaacta gggtggagac
     2461 cgccaccgag accgcctgga tctccttggt gaccgctctg catctagtgc tgggcctcag
     2521 cgccgtcctg ggcctgctgc tgctgaggtg gcagtttcct gcacactaca ggagactgag
     2581 gcatgccctg tggccctcac ttccagacct gcaccgggtc ctaggccagt accttaggga
     2641 cactgcagcc ctgagcccgc ccaaggccac agtctcagat acctgtgaag aagtggaacc
     2701 cagcctcctt gaaatcctcc ccaagtcctc agagaggact cctttgcccc tgtgttcctc
     2761 ccaggcccag atggactacc gaagattgca gccttcttgc ctggggacca tgcccctgtc
     2821 tgtgtgccca cccatggctg agtcagggtc ctgctgtacc acccacattg ccaaccattc
     2881 ctacctacca ctaagctatt ggcagcagcc ttgactcgag cgggatcaat tccgcccccc
     2941 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3001 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3061 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3121 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3181 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3241 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3301 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3361 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     3421 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3481 aaaaacacga taataccatg gcctcctccg aggacgtcat caaggagttc atgcgcttca
     3541 aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     3601 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
     3661 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc
     3721 accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc
     3781 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     3841 acggctcctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc gacggccccg
     3901 taatgcagaa gaagactatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     3961 gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa ggacggcggc cactacctgg
     4021 tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc tactactacg
     4081 tggactccaa gctggacatc acctcccaca acgaggacta caccatcgtg gagcagtacg
     4141 agcgcgccga gggccgccac cacctgttcc tgtagagcgg ccgcgactct agagtcgacc
     4201 tgcaggcatg caagcttcag gtagccggct aacgttaaca accggtacct ctagaactat
     4261 agctagcatg cgcaaattta aagcgctgat atcgataaaa taaaagattt tatttagtct
     4321 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     4381 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     4441 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4501 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     4561 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     4621 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     4681 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     4741 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     4801 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag
     4861 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac
     4921 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga
     4981 cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac
     5041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5821 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7141 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7201 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     7261 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     7321 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     7381 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     7441 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     7501 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     7561 acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca
     7621 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     7681 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     7741 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     7801 ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc
     7861 aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa aagattttat
     7921 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     7981 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     8041 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     8101 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     8161 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     8221 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     8281 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     8341 gcccgggtac ccgt
//
