LOCUS       sequence-359407-        6464 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 6464)
  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 6464)
  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 S248A/T258A
FEATURES             Location/Qualifiers
     source          1..6464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    671..805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        819..1198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1640..1662
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1724..1743
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1771..1780
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1780..1803
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2796..3369
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2819..3371
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2963..2980)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3190..3209
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3371..3769
                     /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(3789..3822)
                     /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(3899..3916)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3905..4129
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4176..4192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4176..4192)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4189..4211)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4200..4216
                     /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(4224..4230,4231..4248,4249..4254))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4269..4290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4349..4544
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4395..4530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4550..4684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4752..4769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4923..5511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5003..5022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5682..6464,1..9,10..78))
                     /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     6305..6324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
       61 gaaatgttga atactcatac tcttcctttt tcaatgaccg ctttggaaaa acaaagactg
      121 tatttcctgg aaattaatgt ttattcaata aactgtgtat tcagctatat tcacatagtg
      181 gtgaggctga aatgaggcgg gaagaggcgg ttggggctta attatatcaa tttgggtggc
      241 cccacagcgc ctccaaggcg ccagtcctgt tttgacaagt tgcctctgga agcctctctt
      301 ctttttctcc agagtaagcg gaggccaggg gcccccggcc tctgcttaat actaaaaaaa
      361 acagctgttg tcatagtaat gattgggtgg aaacattcca ggcctgggtg gagaggcttt
      421 ttgcttcctc ttgcaaaacc acactgacat tccaggcctg ggtggagagg ctttttgctt
      481 cctcttgcaa aaccacactg ccctctggag ggcagttgcc tagcaactaa ttaaaagagg
      541 atgtcgcacg gccagctgcg gtcagttagt cacttcctgc ttaactgact tgacattttc
      601 tattttaaga gtcgggagga aaattactgt gttggaggcc ctccgccatc ttctgaagct
      661 gaatcgaatt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      721 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
      781 caatgtatct tatcatgtct ggatctgata tcatcgtcga cattgattat tgactagtta
      841 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      901 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      961 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     1021 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     1081 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     1141 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1201 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
     1261 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg
     1321 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     1381 cgcagccatt gccttttatg gtaatcgtgc gagagagcgc agggacttcc tttgtcccaa
     1441 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1501 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1561 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1621 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1681 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1741 gtgctgtctc atcattttgg caaagaatta gccaccatgg attacaagga tgacgacgat
     1801 aagctcgaga ccatgtataa catgatggag acggagctga agccgccggg cccgcagcaa
     1861 gcttcggggg gcggcggcgg aggaggcaac gccacggcgg cggcgaccgg cggcaaccag
     1921 aagaacagcc cggaccgcgt caagaggccc atgaacgcct tcatggtatg gtcccggggg
     1981 cagcggcgta agatggccca ggagaacccc aagatgcaca actcggagat cagcaagcgc
     2041 ctgggcgcgg agtggaaact tttgtccgag accgagaagc ggccgttcat cgacgaggcc
     2101 aagcggctgc gcgctctgca catgaaggag cacccggatt ataaataccg gccgcggcgg
     2161 aaaaccaaga cgctcatgaa gaaggataag tacacgcttc ccggaggctt gctggccccc
     2221 ggcgggaaca gcatggcgag cggggttggg gtgggcgccg gcctgggtgc gggcgtgaac
     2281 cagcgcatgg acagctacgc gcacatgaac ggctggagca acggcagcta cagcatgatg
     2341 caggagcagc tgggctaccc gcagcacccg ggcctcaacg ctcacggcgc ggcacagatg
     2401 caaccgatgc accgctacga cgtcagcgcc ctgcagtaca actccatgac cagctcgcag
     2461 acctacatga acggctcgcc cacctacagc atgtcctact cgcagcaggg cacccccggt
     2521 atggcgctgg gctccatggg ctctgtggtc aaggccgagg ccagctccag cccccccgtg
     2581 gttgcctctt cctcccactc cagggcgccc tgccaggccg gggacctccg ggacatgatc
     2641 agcatgtacc tccccggcgc cgaggtgccg gagcccgctg cgcccagtag actgcacatg
     2701 gcccagcact accagagcgg cccggtgccc ggcacggcca ttaacggcac actgcccctg
     2761 tcgcacatgt gagcggccgc gaattccgcc ccccccccct ctccctcccc cccccctaac
     2821 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     2881 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     2941 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     3001 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     3061 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     3121 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     3181 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     3241 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     3301 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3361 cgatgataat atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     3421 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     3481 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     3541 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     3601 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     3661 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     3721 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag aattcgtcga
     3781 gggacccaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
     3841 gttccactag gtacaattcg atatcaagct tatcgatgat ctagagctcg ctgatcagcc
     3901 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3961 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4021 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4081 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4141 gaaagaacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4201 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4261 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4321 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     4381 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4441 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4501 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     4561 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4621 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4681 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4741 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4921 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4981 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5041 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5101 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5161 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5221 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5281 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5341 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5401 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5461 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5521 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5581 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5641 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5701 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5761 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5821 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5881 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5941 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6001 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6061 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6121 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6181 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6241 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6301 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6361 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6421 cgtgcaccca actgatcttc agcatctttt actttcacca gcgt
//
