LOCUS       sequence-410091-        7353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mOct4 S229A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7353)
  TITLE     Hyonchol Jang Lab Oct4 plasmids 2023
REFERENCE   2  (bases 1 to 7353)
  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"
COMMENT     Sequence Label: pMSCV-FLAG-puro-mOct4 S229A
FEATURES             Location/Qualifiers
     source          1..7353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1417..1426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1426..1449
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        2528..3027
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2563..2586)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2960..2979
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3048..3647
                     /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(3048..3067)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3544..3564
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3747..4261
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4423..4439
                     /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(4447..4453,4454..4471,4472..4477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4492..4513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4630..4647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4801..5389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4881..4900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5560..6351,6352..6420))
                     /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     6183..6202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6421..6525)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6593..6611
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6649..6671)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6771..6790
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6984..7006
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7134..7153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7353,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctgcca ccatggatta caaggatgac
     1441 gacgataagc tcgagaccat ggctggacac ctggcttcag acttcgcctt ctcaccccca
     1501 ccaggtgggg gtgatgggtc agcagggctg gagccgggct gggtggatcc tcgaacctgg
     1561 ctaagcttcc aagggcctcc aggtgggcct ggaatcggac caggctcaga ggtattgggg
     1621 atctccccat gtccgcccgc atacgagttc tgcggaggga tggcatactg tggacctcag
     1681 gttggactgg gcctagtccc ccaagttggc gtggagactt tgcagcctga gggccaggca
     1741 ggagcacgag tggaaagcaa ctcagaggga acctcctctg agccctgtgc cgaccgcccc
     1801 aatgccgtga agttggagaa ggtggaacca actcccgagg agtcccagga catgaaagcc
     1861 ctgcagaagg agctagaaca gtttgccaag ctgctgaagc agaagaggat caccttgggg
     1921 tacacccagg ccgacgtggg gctcaccctg ggcgttctct ttggaaaggt gttcagccag
     1981 accaccatct gtcgcttcga ggccttgcag ctcagcctta agaacatgtg taagctgcgg
     2041 cccctgctgg agaagtgggt ggaggaagcc gacaacaatg agaaccttca ggagatatgc
     2101 aaatcggaga ccctggtgca ggcccggaag agaaagcgaa ctgccattga gaaccgtgtg
     2161 aggtggagtc tggagaccat gtttctgaag tgcccgaagc cctccctaca gcagatcact
     2221 cacatcgcca atcagcttgg gctagagaag gatgtggttc gagtatggtt ctgtaaccgg
     2281 cgccagaagg gcaaaagatc aagtattgag tattcccaac gagaagagta tgaggctaca
     2341 gggacacctt tcccaggggg ggctgtatcc tttcctctgc ccccaggtcc ccactttggc
     2401 accccaggct atggaagccc ccacttcacc acactctact cagtcccttt tcctgagggc
     2461 gaggcctttc cctctgttcc cgtcactgct ctgggctctc ccatgcattc aaactgagaa
     2521 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     2581 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     2641 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     2701 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     2761 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     2821 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     2881 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     2941 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     3001 tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca
     3061 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3121 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3181 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3241 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3301 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3361 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     3421 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     3481 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     3541 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     3601 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     3661 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa taaaagattt
     3721 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     3781 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     3841 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     3901 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     3961 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     4021 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     4081 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     4141 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     4201 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     4261 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag
     4321 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt
     4381 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     4441 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6541 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6601 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6661 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6721 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6781 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6841 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6901 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6961 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
     7021 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     7081 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     7141 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     7201 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
     7261 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     7321 aagattttat ttagtctcca gaaaaagggg gga
//
