LOCUS       sequence-153174-        7278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of PU.1 and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7278)
  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 7278)
  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: PU.1-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..7278
                     /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    1010..1577
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1027..1579
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1171..1188)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1398..1417
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1600..1609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1606..2316
                     /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(1751..1769)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2019..2038
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             2365..2879
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3048..3064)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3048..3064)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3061..3083)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3072..3088
                     /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(3096..3102,3103..3120,3121..3126))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3141..3162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3279..3296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3450..4038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3530..3549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4209..5000,5001..5069))
                     /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     4832..4851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5070..5174)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5242..5260
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5298..5320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5420..5439
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5633..5655
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5783..5802
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6002..6518
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6582..6923
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6990..7278,1..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 cacatgttac aggcgtgcaa aatggaaggg ttttccctca
      181 ccgcccctcc atcggatgac ttggttactt acgattcaga gctataccaa cgtccaatgc
      241 atgactacta ctccttcgtg ggcagcgatg gagaaagcca tagcgatcac tactgggatt
      301 tctccgcaca ccatgtccac aacaacgagt ttgagaactt ccctgagaac cacttcacag
      361 agctgcagag cgtgcagccc ccgcagctac agcagctcta tcgccacatg gagctggaac
      421 agatgcacgt cctcgatact cccatggtgc caccccacac cggcctcagt caccaggttt
      481 cctacatgcc ccggatgtgc ttcccttatc aaaccttgtc cccagcccac cagcagagct
      541 cagatgagga ggagggtgag aggcagagcc ctcccctgga ggtgtctgat ggagaagctg
      601 atggcttgga gcctgggcca ggtcttctgc acggggagac aggcagcaag aaaaagattc
      661 gcctgtacca gttcctgctg gacctgctgc gcagcggcga catgaaggac agcatctggt
      721 gggtggacaa ggacaaaggt accttccagt tctcgtccaa gcacaaggag gcgctggcgc
      781 accgctgggg catccagaag ggcaaccgca agaagatgac ctaccagaag atggcgcgcg
      841 cgctgcgcaa ctacggcaag acaggcgagg tgaagaaagt caagaagaag ctcacctacc
      901 agttcagcgg cgaggtgctg ggccgtgggg gcctggccga gcggcgcctc ccgccccact
      961 gatcgcggcc gctacgtaaa ttgaattctg ctatacgaag ttatccctct ccctcccccc
     1021 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     1081 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     1141 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     1201 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     1261 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     1321 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     1381 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     1441 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc tttacatgtg
     1501 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     1561 gaagaacacg atgataatat ggccacaacc atgactagtg ccaccatggt gagcaagggc
     1621 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     1681 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1741 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     1801 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     1861 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1921 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1981 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2041 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2101 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2161 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2221 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2281 cactccaccg gcggcatgga cgagctgtac aagtaagcat cgataaaata aaagatttta
     2341 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     2401 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     2461 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     2521 ttcctgcccc cggctcaggg ccaagaacag atggtccccg atgcggtccc gccctcagca
     2581 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     2641 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     2701 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     2761 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     2821 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     2881 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     2941 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     3001 attatggaca gcgcagaaga gctggggaga attaattcgt aatcatggtc atagctgttt
     3061 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3121 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3181 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3241 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3301 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3361 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4021 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4081 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4141 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4201 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4261 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4321 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4381 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4441 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4501 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4561 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4621 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4681 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4741 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4801 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4861 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4921 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4981 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5041 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5101 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5161 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5221 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5281 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5341 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5401 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5461 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     5521 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5581 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     5641 cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     5701 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     5761 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     5821 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta
     5881 aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc
     5941 ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg
     6001 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     6061 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     6121 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     6181 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     6241 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     6301 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     6361 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
     6421 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
     6481 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
     6541 agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
     6601 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
     6661 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     6721 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     6781 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     6841 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     6901 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     6961 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     7021 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     7081 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     7141 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     7201 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     7261 acccccctcc ctgggtca
//
