LOCUS       sequence-432115-        8142 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Overexpress human SOX2 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 8142)
  TITLE     Jauch lab plasmids
REFERENCE   2  (bases 1 to 8142)
  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: pHAGE2-Ef1a-SOX2-3xFLAG
FEATURES             Location/Qualifiers
     source          1..8142
                     /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             4499..4564
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4584..5172
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4637..4657)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5055..5066)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5320..5500
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5547..5565)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5633..5737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5738..5806,5807..6598)
                     /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(5956..5975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6769..7357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7511..7528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7768..7789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7804..7809,7810..7827,7828..7834)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7842..7858
                     /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     7847..7869
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7866..7882
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7866..7882
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7932..7951
                     /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 tgtacaacat
     3541 gatggagacg gagctgaagc cgccgggccc gcagcaaact tcggggggcg gcggcggcaa
     3601 ctccaccgcg gcggcggccg gcggcaacca gaaaaacagc ccggaccgcg tcaagcggcc
     3661 catgaatgcc ttcatggtgt ggtcccgcgg gcagcggcgc aagatggccc aagagaaccc
     3721 caagatgcac aactcggaga tcagcaagcg cctgggcgcc gagtggaaac ttttgtcgga
     3781 gacggagaag cggccgttca tcgacgaggc taagcggctg cgagcgctgc acatgaagga
     3841 gcacccggat tataaatacc ggccccggcg gaaaaccaag acgctcatga agaaggataa
     3901 gtacacgctg cccggcgggc tgctggcccc cggcggcaat agcatggcga gcggggtcgg
     3961 ggtgggcgcc ggcctgggcg cgggcgtgaa ccagcgcatg gacagttacg cgcacatgaa
     4021 cggctggagc aacggcagct acagcatgat gcaggaccag ctgggctacc cgcagcaccc
     4081 gggcctcaat gcgcacggcg cagcgcagat gcagcccatg caccgctacg acgtgagcgc
     4141 cctgcagtac aactccatga ccagctcgca gacctacatg aacggctcgc ccacctacag
     4201 catgtcctac tcgcagcagg gcacccctgg catggctctt ggctccatgg gttcggtggt
     4261 caagtccgag gccagctcca gcccccctgt ggttacctct tcctcccact ccagggcgcc
     4321 ctgccaggcc ggggacctcc gggacatgat cagcatgtat ctccccggcg ccgaggtgcc
     4381 ggaacccgcc gcccccagca gacttcacat gtcccagcac taccagagcg gcccggtgcc
     4441 cggcacggcc attaacggca cactgcccct ctcacacatg attacgcgtg gcagcggcga
     4501 ctacaaggac cacgacggcg actacaagga ccacgacatc gactacaagg acgacgacga
     4561 caagtagtaa atcgatagat cctaatcaac ctctggatta caaaatttgt gaaagattga
     4621 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4681 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4741 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4801 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4861 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4921 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4981 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5041 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5101 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5161 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5221 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5281 cacctgcagg acaggcgcgc cctgcttttt gcttgtactg ggtctctctg gttagaccag
     5341 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     5401 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     5461 tccctcagac ccttttagtc agtgtggaaa atctctagca cccgggcgat taaggaaagg
     5521 gctagatcat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5581 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5641 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5701 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5761 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5821 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5881 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5941 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     6001 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6061 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6121 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6181 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6241 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6301 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6361 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6421 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6481 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6541 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6601 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6661 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6721 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6781 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6841 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6901 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6961 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7021 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7081 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7141 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7201 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7261 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7321 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7381 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7441 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7501 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7561 tctccccgcg cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt
     7621 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     7681 ttttttggag gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg
     7741 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7801 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7861 tcacacagga aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact
     7921 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
     7981 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
     8041 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
     8101 aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
