LOCUS       sequence-256120-        7850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of target proteins BAP-Sox2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7850)
  AUTHORS   Kulyyassov A, Ogryzko V
  TITLE     In Vivo Quantitative Estimation of DNA-Dependent Interaction of Sox2
            and Oct4 Using BirA-Catalyzed Site-Specific Biotinylation.
  JOURNAL   Biomolecules. 2020 Jan 16;10(1). pii: biom10010142. doi: 
            10.3390/biom10010142.
  PUBMED    31963153
REFERENCE   2  (bases 1 to 7850)
  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: 
            "Biomolecules."; date: "2020-01-16"; volume: "10(1). pii: 
            biom10010142. doi"; pages: " 10.3390/biom10010142"
COMMENT     Sequence Label: pcDNA3-BAP-Sox2
FEATURES             Location/Qualifiers
     source          1..7850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..48)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    37..261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      307..735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(394..413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     604..625
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(744..764)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        749..1078
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      929..1064
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     991..1010
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1145..1939
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1199..1218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1809..1828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2113..2234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2150..2169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2204..2223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2283..2299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2283..2299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2296..2318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2307..2323
                     /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(2331..2337,2338..2355,2356..2361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2376..2397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2514..2531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2685..3270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2765..2784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3441..4232,4233..4301))
                     /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     4064..4083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4302..4406)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4481..4500)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4672..5051
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5052..5255
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5206..5226
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5300..5319
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5300..5318
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5384..5404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     misc_feature    6484..7033
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6628..6645)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6855..6874
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
       61 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
      121 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
      181 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
      241 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
      301 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      361 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      421 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      481 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      541 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      601 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      661 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      721 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      781 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      841 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      901 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      961 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     1021 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     1081 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     1141 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1201 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1261 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1321 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1381 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1441 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1501 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1561 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1621 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1681 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1741 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1801 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1861 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1921 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     1981 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     2041 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     2101 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2161 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2221 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     2281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2341 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2521 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2581 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2641 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2701 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2761 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2821 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2881 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2941 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3001 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3061 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3121 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3181 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     3241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     4441 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     4501 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     4561 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     4621 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4681 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4741 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4801 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4861 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4921 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4981 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     5041 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     5101 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5161 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5221 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     5281 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
     5341 taaacttaag cttggtaccg agctcggatc ttgaaccatg ggacaccatc accatcacca
     5401 tcatggcctg acaagaatcc tggaagctca gaagatcgtg agaggaggcc tcgagggcat
     5461 gtacaacatg atggagacgg agctgaagcc gccgggcccg cagcaaactt cggggggcgg
     5521 cggcggcaac tccaccgcgg cggcggccgg cggcaaccag aaaaacagcc cggaccgcgt
     5581 caagcggccc atgaatgcct tcatggtgtg gtcccgcggg cagcggcgca agatggccca
     5641 ggagaacccc aagatgcaca actcggagat cagcaagcgc ctgggcgccg agtggaaact
     5701 tttgtcggag acggagaagc ggccgttcat cgacgaggct aagcggctgc gagcgctgca
     5761 catgaaggag cacccggatt ataaataccg gccccggcgg aaaaccaaga cgctcatgaa
     5821 gaaggataag tacacgctgc ccggcgggct gctggccccc ggcggcaata gcatggcgag
     5881 cggggtcggg gtgggcgccg gcctgggcgc gggcgtgaac cagcgcatgg acagttacgc
     5941 gcacatgaac ggctggagca acggcagcta cagcatgatg caggaccagc tgggctaccc
     6001 gcagcacccg ggcctcaatg cgcacggcgc agcgcagatg cagcccatgc accgctacga
     6061 cgtgagcgcc ctgcagtaca actccatgac cagctcgcag acctacatga acggctcgcc
     6121 cacctacagc atgtcctact cgcagcaggg cacccctggc atggctcttg gctccatggg
     6181 ttcggtggtc aagtccgagg ccagctccag cccccctgtg gttacctctt cctcccactc
     6241 cagggcgccc tgccaggccg gggacctccg ggacatgatc agcatgtatc tccccggcgc
     6301 cgaggtgccg gaacccgccg cccccagcag acttcacatg tcccagcact accagagcgg
     6361 cccggtgccc ggcacggcca ttaacggcac actgcccctc tcacacatgt gagcggccgc
     6421 atcgagagat ccgggagaca caacggtttc cctctagcgg gatcaattcc gccccccccc
     6481 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     6541 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     6601 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     6661 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     6721 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     6781 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     6841 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     6901 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     6961 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     7021 aaacacgata ataccatgga ttcatacctg ctgatgtggg gactgctcac gttcatcatg
     7081 gtgcctggct gccaggcaga gctctgtgac gatgacccgc cagagatccc acacgccaca
     7141 ttcaaagcca tggcctacaa ggaaggaacc atgttgaact gtgaatgcaa gagaggtttc
     7201 cgcagaataa aaagcgggtc actctatatg ctctgtacag gaaactctag ccactcgtcc
     7261 tgggacaacc aatgtcaatg cacaagctct gccactcgga acacaacgaa acaagtgaca
     7321 cctcaacctg aagaacagaa agaaaggaaa accacagaaa tgcaaagtcc aatgcagcca
     7381 gtggaccaag cgagccttcc aggtcactgc agggaacctc caccatggga aaatgaagcc
     7441 acagagagaa tttatcattt cgtggtgggg cagatggttt attatcagtg cgtccaggga
     7501 tacagggctc tacacagagg tcctgctgag agcgtctgca aaatgaccca cgggaagaca
     7561 aggtggaccc agccccagct catatgcaca ggtgaaatgg agaccagtca gtttccaggt
     7621 gaagagaagc ctcaggcaag ccccgaaggc cgtcctgaga gtgagacttc ctgcctcgtc
     7681 acaacaacag attttcaaat acagacagaa atggctgcaa ccatggagac gtccatattt
     7741 acaacagagt accaggtagc agtggccggc tgtgttttcc tgctgatcag cgtcctcctc
     7801 ctgagtgggc tcacctggca gcggagacag aggaagagta gaagaacaat
//
