LOCUS       sequence-410097-        7353 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of mOct4 T87A 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 Apr 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 T87A
FEATURES             Location/Qualifiers
     source          1..7353
                     /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"
     promoter        2527..3026
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2562..2585)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2959..2978
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3047..3646
                     /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(3047..3066)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3543..3563
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3746..4260
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4422..4438
                     /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(4446..4452,4453..4470,4471..4476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4491..4512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4629..4646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4880..4899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5559..6350,6351..6419))
                     /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     6182..6201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6420..6524)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6592..6610
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6648..6670)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6770..6789
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6983..7005
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7133..7152
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7352..7353,1..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 tggaggcatt 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 catgcattca aactgagaat
     2521 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     2581 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     2641 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     2701 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     2761 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     2821 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     2881 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     2941 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     3001 cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga ccgagtacaa
     3061 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3121 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3181 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3241 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3301 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3361 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3421 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3481 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3541 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3601 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     3661 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt
     3721 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     3781 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     3841 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     3901 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     3961 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4021 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4081 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     4141 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4201 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     4261 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     4321 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     4381 cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt
     4441 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4501 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4561 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6541 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6601 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6661 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6721 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     6781 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     6841 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6901 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     6961 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg
     7021 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     7081 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     7141 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     7201 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     7261 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     7321 agattttatt tagtctccag aaaaaggggg gaa
//
