LOCUS       sequence-153214-        6751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of PU.1-ets (competitive inhibitor of PU.1) 
            and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6751)
  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 6751)
  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-ets-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..6751
                     /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    483..1050
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    500..1052
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(644..661)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     871..890
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1073..1082
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1079..1789
                     /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(1224..1242)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1492..1511
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             1838..2352
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2521..2537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2521..2537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2534..2556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2545..2561
                     /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(2569..2575,2576..2593,2594..2599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2614..2635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2752..2769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2923..3511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3003..3022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3682..4473,4474..4542))
                     /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     4305..4324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4543..4647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4715..4733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4771..4793)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4893..4912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5106..5128
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5256..5275
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5475..5991
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6055..6396
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6463..6751,1..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 atgccaggtc ttctgcacgg ggagacaggc agcaagaaaa
      181 agattcgcct gtaccagttc ctgctggacc tgctgcgcag cggcgacatg aaggacagca
      241 tctggtgggt ggacaaggac aaaggtacct tccagttctc gtccaagcac aaggaggcgc
      301 tggcgcaccg ctggggcatc cagaagggca accgcaagaa gatgacctac cagaagatgg
      361 cgcgcgcgct gcgcaactac ggcaagacag gcgaggtgaa gaaagtcaag aagaagctca
      421 cctaccagtt cagcggcgag gtgctgggcc gttgagaatt ctgctatacg aagttatccc
      481 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
      541 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
      601 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
      661 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
      721 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
      781 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
      841 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
      901 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca
      961 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     1021 gtggttttcc tttgaagaac acgatgataa tatggccaca accatgacta gtgccaccat
     1081 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1141 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1201 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1261 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     1321 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     1381 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1441 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     1501 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     1561 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     1621 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     1681 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     1741 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag catcgataaa
     1801 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     1861 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     1921 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
     1981 ctgtggtaag cagttcctgc ccccggctca gggccaagaa cagatggtcc ccgatgcggt
     2041 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     2101 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     2161 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     2221 cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     2281 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     2341 ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt
     2401 cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact
     2461 cctgaaatag ttcattatgg acagcgcaga agagctgggg agaattaatt cgtaatcatg
     2521 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2581 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2641 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2701 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2761 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2821 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2881 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2941 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3001 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3061 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3121 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3181 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3241 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3301 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3361 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3421 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3481 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3541 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3601 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3661 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3721 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3781 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3841 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3901 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3961 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4021 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4081 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4141 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4201 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4261 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4321 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4381 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4441 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4501 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4561 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4621 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4681 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4741 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4801 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4861 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4921 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     4981 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5041 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5101 gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg
     5161 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     5221 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     5281 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta
     5341 gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat
     5401 caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc
     5461 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     5521 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     5581 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5641 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     5701 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     5761 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     5821 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     5881 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct
     5941 tgggagggtc tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc
     6001 caccgagatt tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg
     6061 gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt
     6121 ttgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
     6181 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
     6241 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
     6301 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
     6361 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
     6421 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
     6481 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     6541 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     6601 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     6661 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
     6721 gccttggctt ttgacccccc tccctgggtc a
//
