LOCUS       sequence-432004-        8271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpress human OCT4 in HEK 293T cells for use in whole cell 
            extract EMSAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8271)
  AUTHORS   Ho SY, Hu H, Ho DHH, Renom APS, Yeung SW, Boerner F, Weng M, 
            Hutchins AP, Jauch R
  TITLE     An acidic residue within the OCT4 dimerization interface of SOX17 is
            necessary and sufficient to overcome its pluripotency-inducing 
            activity.
  JOURNAL   Stem Cell Reports. 2025 Mar 11;20(3):102398. doi: 
            10.1016/j.stemcr.2025.102398. Epub 2025 Feb 6.
  PUBMED    39919754
REFERENCE   2  (bases 1 to 8271)
  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"; journalName: "Stem Cell 
            Reports."; date: "2025-03-11"; volume: "20(3):102398. doi"; pages: "
            10.1016/j.stemcr.2025.102398. Epub 2025 Feb 6"
COMMENT     Sequence Label: pHAGE2-Ef1a-OCT4-3xFLAG
FEATURES             Location/Qualifiers
     source          1..8271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2307..3485
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2538..3476
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3433..3453
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4628..4693
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4713..5301
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4766..4786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5184..5195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5449..5629
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5676..5694)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5762..5866
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5867..5935,5936..6727)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6085..6104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6898..7486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7640..7657
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7897..7918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7933..7938,7939..7956,7957..7963)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7971..7987
                     /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)."
     primer_bind     7976..7998
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7995..8011
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7995..8011
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8061..8080
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgat
     2221 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     2281 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     2341 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     2401 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2461 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2521 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     2581 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     2641 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     2701 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     2761 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     2821 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     2881 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     2941 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     3001 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     3061 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     3121 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     3181 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     3241 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     3301 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     3361 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     3421 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     3481 cgtgagcggc caagggcgaa ttctgcagat atccatcaca ctggctagca tggcgggaca
     3541 cctggcttcg gatttcgcct tctcgccccc tccaggtggt ggaggtgatg ggccaggggg
     3601 gccggagccg ggctgggttg atcctcggac ctggctaagc ttccaaggcc ctcctggagg
     3661 gccaggaatc gggccggggg ttgggccagg ctctgaggtg tgggggattc ccccatgccc
     3721 cccgccgtat gagttctgtg gggggatggc gtactgtggg ccccaggttg gagtggggct
     3781 agtgccccaa ggcggcttgg agacctctca gcctgagggc gaagcaggag tcggggtgga
     3841 gagcaactcc gatggggcct ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct
     3901 ggagaaggag aagctggagc aaaacccgga ggagtcccag gacatcaaag ctctgcagaa
     3961 agaactcgag caatttgcca agctcctgaa gcagaagagg atcaccctgg gatatacaca
     4021 ggccgatgtg gggctcaccc tgggggttct atttgggaag gtattcagcc aaacgaccat
     4081 ctgccgcttt gaggctctgc agcttagctt caagaacatg tgtaagctgc ggcccttgct
     4141 gcagaagtgg gtggaggaag ctgacaacaa tgaaaatctt caggagatat gcaaagcaga
     4201 aaccctcgtg caggcccgaa agagaaagcg aaccagtatc gagaaccgag tgagaggcaa
     4261 cctggagaat ttgttcctgc agtgcccgaa acccacactg cagcagatca gccacatcgc
     4321 ccagcagctt gggctcgaga aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa
     4381 gggcaagcga tcaagcagcg actatgcaca acgagaggat tttgaggctg ctgggtctcc
     4441 tttctcaggg ggaccagtgt cctttcctct ggccccaggg ccccattttg gtaccccagg
     4501 ctatgggagc cctcacttca ctgcactgta ctcctcggtc cctttccctg agggggaagc
     4561 ctttccccct gtctccgtca ccactctggg ctctcccatg cattcaaaca ttacgcgtgg
     4621 cagcggcgac tacaaggacc acgacggcga ctacaaggac cacgacatcg actacaagga
     4681 cgacgacgac aagtagtaaa tcgatagatc ctaatcaacc tctggattac aaaatttgtg
     4741 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     4801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5101 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5281 ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta
     5341 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5401 agacaagatc acctgcagga caggcgcgcc ctgctttttg cttgtactgg gtctctctgg
     5461 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5521 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5581 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcac ccgggcgatt
     5641 aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     5701 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5761 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5821 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5881 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5941 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6001 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6061 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     6121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6241 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6421 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6481 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6721 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7621 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7681 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca tggctgacta
     7741 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     7801 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc acgacaggtt
     7861 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     7921 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     7981 ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa ataaagcatt
     8041 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     8101 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
     8161 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
     8221 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t
//
