LOCUS       sequence-359405-        7242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Sox2 mutants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  AUTHORS   Kim DK, Lee JS, Lee EY, Jang H, Han S, Kim HY, Hwang IY, Choi JW, 
            Shin HM, You HJ, Youn HD, Jang H
  TITLE     O-GlcNAcylation of Sox2 at threonine 258 regulates the self-renewal 
            and early cell fate of embryonic stem cells.
  JOURNAL   Exp Mol Med. 2021 Nov;53(11):1759-1768. doi: 
            10.1038/s12276-021-00707-7. Epub 2021 Nov 24.
  PUBMED    34819616
REFERENCE   2  (bases 1 to 7242)
  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: 
            "10.1038/s12276-021-00707-7"; journalName: "Exp Mol Med"; date: 
            "2021-11"; volume: "53"; issue: "11"; pages: "1759-1768"
COMMENT     Sequence Label: pCAG-FLAG-IRES-blasticidin-Sox2 T258A
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    576..710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        724..1103
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1383..2399
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2323..2345
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2407..2426
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2454..2463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2463..2486
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3479..4052
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3502..4054
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3646..3663)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3873..3892
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4054..4452
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(4472..4505)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4582..4599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4588..4812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4859..4875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4859..4875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4872..4894)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4883..4899
                     /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(4907..4913,4914..4931,4932..4937))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4952..4973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5032..5227
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5078..5213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5233..5367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5435..5452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5606..6194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5686..5705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6365..7156,7157..7225))
                     /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     6988..7007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccgctttg gaaaaacaaa gactgtattt cctggaaatt aatgtttatt caataaactg
       61 tgtattcagc tatattcaca tagtggtgag gctgaaatga ggcgggaaga ggcggttggg
      121 gcttaattat atcaatttgg gtggccccac agcgcctcca aggcgccagt cctgttttga
      181 caagttgcct ctggaagcct ctcttctttt tctccagagt aagcggaggc caggggcccc
      241 cggcctctgc ttaatactaa aaaaaacagc tgttgtcata gtaatgattg ggtggaaaca
      301 ttccaggcct gggtggagag gctttttgct tcctcttgca aaaccacact gacattccag
      361 gcctgggtgg agaggctttt tgcttcctct tgcaaaacca cactgccctc tggagggcag
      421 ttgcctagca actaattaaa agaggatgtc gcacggccag ctgcggtcag ttagtcactt
      481 cctgcttaac tgacttgaca ttttctattt taagagtcgg gaggaaaatt actgtgttgg
      541 aggccctccg ccatcttctg aagctgaatc gaattaactt gtttattgca gcttataatg
      601 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
      661 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatatcatc
      721 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      781 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      841 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      901 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      961 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     1021 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1081 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     1141 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     1201 cgatgggggc gggggggggg ggggggcgcg cgccrggggg gggggggggg ggsgrggggs
     1261 ggggsggggs grggsggrgr ggkgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     1321 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     1381 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     1441 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     1501 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
     1561 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
     1621 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
     1681 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1741 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1801 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1861 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1921 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1981 cggggccgcc tcgggccggg grgggctcgg gggrggggcg cggcggcccc cggagcgccg
     2041 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     2101 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     2161 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     2221 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     2281 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     2341 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     2401 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttagccacca
     2461 tggattacaa ggatgacgac gataagctcg agaccatgta taacatgatg gagacggagc
     2521 tgaagccgcc gggcccgcag caagcttcgg ggggcggcgg cggaggaggc aacgccacgg
     2581 cggcggcgac cggcggcaac cagaagaaca gcccggaccg cgtcaagagg cccatgaacg
     2641 ccttcatggt atggtcccgg gggcagcggc gtaagatggc ccaggagaac cccaagatgc
     2701 acaactcgga gatcagcaag cgcctgggcg cggagtggaa acttttgtcc gagaccgaga
     2761 agcggccgtt catcgacgag gccaagcggc tgcgcgctct gcacatgaag gagcacccgg
     2821 attataaata ccggccgcgg cggaaaacca agacgctcat gaagaaggat aagtacacgc
     2881 ttcccggagg cttgctggcc cccggcggga acagcatggc gagcggggtt ggggtgggcg
     2941 ccggcctggg tgcgggcgtg aaccagcgca tggacagcta cgcgcacatg aacggctgga
     3001 gcaacggcag ctacagcatg atgcaggagc agctgggcta cccgcagcac ccgggcctca
     3061 acgctcacgg cgcggcacag atgcaaccga tgcaccgcta cgacgtcagc gccctgcagt
     3121 acaactccat gaccagctcg cagacctaca tgaacggctc gcccacctac agcatgtcct
     3181 actcgcagca gggcaccccc ggtatggcgc tgggctccat gggctctgtg gtcaagtccg
     3241 aggccagctc cagccccccc gtggttgcct cttcctccca ctccagggcg ccctgccagg
     3301 ccggggacct ccgggacatg atcagcatgt acctccccgg cgccgaggtg ccggagcccg
     3361 ctgcgcccag tagactgcac atggcccagc actaccagag cggcccggtg cccggcacgg
     3421 ccattaacgg cacactgccc ctgtcgcaca tgtgagcggc cgcgaattcc gccccccccc
     3481 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     3541 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     3601 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     3661 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     3721 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3781 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     3841 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3901 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     3961 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     4021 acgtggtttt cctttgaaaa acacgatgat aatatggcca agcctttgtc tcaagaagaa
     4081 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     4141 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     4201 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     4261 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     4321 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     4381 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     4441 tgggagggct aagaattcgt cgagggaccc aataacttcg tatagcatac attatacgaa
     4501 gttatacatg tttaagggtt ccggttccac taggtacaat tcgatatcaa gcttatcgat
     4561 gatctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4621 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4681 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4741 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4801 tgggctctat ggcttctgag gcggaaagaa cctgcagccc aagcttggcg taatcatggt
     4861 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4921 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4981 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     5041 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5101 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5161 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5221 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5281 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5341 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7201 cgacacggaa atgttgaata ctcatactct tcctttttca at
//
