LOCUS       sequence-153233-        7049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of p27 and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  AUTHORS   Kueh HY, Champhekar A, Nutt SL, Elowitz MB, Rothenberg EV
  TITLE     Positive feedback between PU.1 and the cell cycle controls myeloid 
            differentiation.
  JOURNAL   Science. 2013 Aug 9;341(6146):670-3. doi: 10.1126/science.1240831. 
            Epub 2013 Jul 18.
  PUBMED    23868921
REFERENCE   2  (bases 1 to 7049)
  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"; doi: 
            "10.1126/science.1240831"; journalName: "Science"; date: "2013-08-9-
            9"; volume: "341"; issue: "6146"; pages: "670-3"
COMMENT     Sequence Label: p27-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    781..1348
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    798..1350
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(942..959)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1169..1188
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1371..1380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1377..2087
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1522..1540)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1790..1809
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             2136..2650
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2819..2835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2819..2835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2832..2854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2843..2859
                     /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(2867..2873,2874..2891,2892..2897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2912..2933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3050..3067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3221..3809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3301..3320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3980..4771,4772..4840))
                     /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     4603..4622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4841..4945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5013..5031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5069..5091)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5191..5210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5404..5426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5554..5573
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5773..6289
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6353..6694
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6761..7049,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct gccaccatgt caaacgtgcg agtgtctaac gggagcccta
      181 gcctggagcg gatggacgcc aggcaggcgg agcaccccaa gccctcggcc tgcaggaacc
      241 tcttcggccc ggtggaccac gaagagttaa cccgggactt ggagaagcac tgcagagaca
      301 tggaagaggc gagccagcgc aagtggaatt tcgattttca gaatcacaaa cccctagagg
      361 gcaagtacga gtggcaagag gtggagaagg gcagcttgcc cgagttctac tacagacccc
      421 cgcggccccc caaaggtgcc tgcaaggtgc cggcgcagga gagccaggat gtcagcggga
      481 gccgcccggc ggcgccttta attggggctc cggctaactc tgaggacacg catttggtgg
      541 acccaaagac tgatccgtcg gacagccaga cggggttagc ggagcaatgc gcaggaataa
      601 ggaagcgacc tgcaaccgac gattcttcta ctcaaaacaa aagagccaac agaacagaag
      661 aaaatgtttc agacggttcc ccaaatgccg gttctgtgga gcagacgccc aagaagcctg
      721 gcctcagaag acgtcaaacg taacggagct taagaattct gctatacgaa gttatccctc
      781 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
      841 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
      901 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
      961 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     1021 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     1081 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     1141 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     1201 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg
     1261 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     1321 ggttttcctt tgaagaacac gatgataata tggccacaac catgactagt gccaccatgg
     1381 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     1441 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1501 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     1561 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     1621 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     1681 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     1741 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1801 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1861 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1921 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1981 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2041 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagca tcgataaaat
     2101 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     2161 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     2221 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     2281 gtggtaagca gttcctgccc ccggctcagg gccaagaaca gatggtcccc gatgcggtcc
     2341 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     2401 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     2461 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     2521 atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     2581 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     2641 ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg
     2701 aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc
     2761 tgaaatagtt cattatggac agcgcagaag agctggggag aattaattcg taatcatggt
     2821 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2881 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2941 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3001 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3061 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3121 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3661 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3721 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3781 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3841 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3901 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3961 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4021 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4081 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4141 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4201 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4261 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4321 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4381 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4441 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4501 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4561 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4621 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4681 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4741 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4801 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4861 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4921 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4981 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5041 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5101 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5161 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5221 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     5281 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5341 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     5401 tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc
     5461 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5521 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5581 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt
     5641 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     5701 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     5761 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5821 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     5881 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     5941 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     6001 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     6061 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     6121 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     6181 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg ctgatccttg
     6241 ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt ggaggttcca
     6301 ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag gtaagctggc
     6361 cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt
     6421 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     6481 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     6541 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     6601 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     6661 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     6721 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     6781 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     6841 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     6901 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     6961 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     7021 cttggctttt gacccccctc cctgggtca
//
