LOCUS       sequence-410080-        7304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mOct4 S351A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7304)
  TITLE     Hyonchol Jang Lab Oct4 plasmids 2023
REFERENCE   2  (bases 1 to 7304)
  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 S351A
FEATURES             Location/Qualifiers
     source          1..7304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1416..1425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1425..1448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2560..2583)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2910..2929
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2998..3597
                     /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(2998..3017)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3494..3514
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3697..4211
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4373..4389
                     /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(4397..4403,4404..4421,4422..4427))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4442..4463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4580..4597)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4751..5339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4831..4850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5510..6301,6302..6370))
                     /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     6133..6152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6371..6475)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6543..6561
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6599..6621)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6721..6740
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6934..6956
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7084..7103
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7303..7304,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctgccac catggattac aaggatgacg
     1441 acgataagct cgagaccatg gctggacacc tggcttcaga cttcgccttc tcacccccac
     1501 caggtggggg tgatgggtca gcagggctgg agccgggctg ggtggatcct cgaacctggc
     1561 taagcttcca agggcctcca ggtgggcctg gaatcggacc aggctcagag gtattgggga
     1621 tctccccatg tccgcccgca tacgagttct gcggagggat ggcatactgt ggacctcagg
     1681 ttggactggg cctagtcccc caagttggcg tggagacttt gcagcctgag ggccaggcag
     1741 gagcacgagt ggaaagcaac tcagagggaa cctcctctga gccctgtgcc gaccgcccca
     1801 atgccgtgaa gttggagaag gtggaaccaa ctcccgagga gtcccaggac atgaaagccc
     1861 tgcagaagga gctagaacag tttgccaagc tgctgaagca gaagaggatc accttggggt
     1921 acacccaggc cgacgtgggg ctcaccctgg gcgttctctt tggaaaggtg ttcagccaga
     1981 ccaccatctg tcgcttcgag gccttgcagc tcagccttaa gaacatgtgt aagctgcggc
     2041 ccctgctgga gaagtgggtg gaggaagccg acaacaatga gaaccttcag gagatatgca
     2101 aatcggagac cctggtgcag gcccggaaga gaaagcgaac tagcattgag aaccgtgtga
     2161 ggtggagtct ggagaccatg tttctgaagt gcccgaagcc ctccctacag cagatcactc
     2221 acatcgccaa tcagcttggg ctagagaagg atgtggttcg agtatggttc tgtaaccggc
     2281 gccagaaggg caaaagatca agtattgagt attcccaacg agaagagtat gaggctacag
     2341 ggacaccttt cccagggggg gctgtatcct ttcctctgcc cccaggtccc cactttggca
     2401 ccccaggcta tggaagcccc cacttcacca cactctactc agtccctttt cctgagggcg
     2461 aggcctttcc ctctgttccc gtcactgctc tgggctctcc catgcatgca aactgaattc
     2521 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     2581 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     2641 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     2701 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     2761 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     2821 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca ggggcggggc
     2881 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     2941 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     3001 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     3061 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     3121 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     3181 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     3241 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     3301 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     3361 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     3421 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     3481 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     3541 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc
     3601 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa
     3661 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3721 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     3781 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     3841 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     3901 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     3961 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     4021 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     4081 gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     4141 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     4201 gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc
     4261 gaatattaag taatcctgac tcaattagcc actgttttga atccacatac tccaatactc
     4321 ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc
     4381 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4441 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4501 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4561 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4621 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4681 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4741 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4801 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4861 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4921 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4981 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5041 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5101 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5161 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5221 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5281 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5341 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5401 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5461 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5521 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5581 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5641 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5701 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5761 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5821 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5881 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5941 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6001 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6061 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6121 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6181 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6241 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6301 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6361 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6421 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6481 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6541 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6601 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6661 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6721 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     6781 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     6841 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6901 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6961 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     7021 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     7081 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     7141 cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt
     7201 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     7261 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaa
//
