LOCUS       sequence-230186-        7359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor TOX2 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7359)
  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"
COMMENT     Sequence Label: pTOX2-donor
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..50)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(47..69)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    58..74
                     /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(82..88,89..106,107..112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(265..282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(436..1024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(516..535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1195..1986,1987..2055))
                     /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     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2228..2246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2284..2306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2406..2425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2619..2641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2633..2650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2634..2650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3891..3911
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3924..3968
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4008..4031
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4059..4061,4062..4064,4065..4778)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4104..4125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4365..4384)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4816..5389,5390..5392,5393..5401)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4982..4999)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5209..5228
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5483..6286
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5546..5565)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6156..6175
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cgactgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
       61 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2221 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2461 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2641 gacggccagt gaattcgagc tcggtaccca agaatgcagg acaggggacc tcggtgggac
     2701 acggtgggag aagggaccca ctgtctatcc caaggaatta cagacacctt agagacccct
     2761 tctccctgca ccccagtgtc ctccctagtc ctcctgtggg cctcctgtct caggcctctg
     2821 ttcctatcct gcactccacc tttcaaaccc atgctaacac ctccctctag aaagtcttcc
     2881 tggatccacg ccaccccaac tctggggaag agtgaccttt caacaaccgc agccagcagg
     2941 agagggagag tgcagtgagg agggacactt gctccttaac ccgccagcca ggttcccatt
     3001 cccttggcaa aaatgagtcc cgcagccaca cctgggggac gtagtgccaa gcagctgtcc
     3061 cagcgatagc gccacaacct ggaaggcaca gcgcacttgt cgggtggcca ccttgctgct
     3121 gagaaacgct gcccccagcc ctagcctcgg cctcccagca cacggctggt tcctctgctg
     3181 acccaccagc gcccttggac actgaacaag tgaccagcgc gcatgcccca tttctgacat
     3241 gcctgaagtg agattctaca tgggtgggtg gactcagctc agcaagaatc ccttccctgc
     3301 accccaagca ggtgcagacc ctcctacccc agccctttcc tgtccctaag tttccttctc
     3361 aaaagtcaga atacttaggg tatgtagggg agtgtcacac aatgtccaag cctcctgagg
     3421 cttgcccatc acatccagca tggaggtcag accccagcag ggccgtattt taaaaacata
     3481 aaaagcacct gtcctccctt aattcagttt gagatttaca aattcctgtg attaaaaacc
     3541 catgtgagag caatggtgac atcggcgtgc gcctcgatga aagacagaca cgctactgcc
     3601 ctttcaccca gagctgagga ccgggtggtc ccctgctggg tttccgggag gggtttggtt
     3661 ctgcttggtg tagaggtggg tgggggtggg acttgcagat gcaagtccct cattcagcca
     3721 cagaggggca gctgaatcca ggggtggggg tggggcgtac agtgcagggg tcctggtgct
     3781 cccctggccc ggagaggtac ccaacagctt tgacagcccc tcctctctct cacagcctgc
     3841 tccccaggga caaatcgctc tacctcaccg ggattccaac tactgcaagc gagaatcttt
     3901 attttcaggg cgccgccaaa ttcaaagaaa ccgctgctgc taaattcgaa cgccagcaca
     3961 tggacagcgg aggtggaggt tcgagcggtc cctcgggttc gtcgagcctg gaagttctgt
     4021 tccaggggcc cctgtcgtcg agcggtccct cgggttcgat ggtgagcaaa ggcgaggagc
     4081 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     4141 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4201 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4261 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4321 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     4381 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4441 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     4501 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     4561 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     4621 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     4681 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct taaggagttc gtgaccgccg
     4741 ccgggatcac tctcggcatg gacgagctgt acaagtaatg atctagagtc gagttaatta
     4801 agaattccgc ccccccccct ctccctcccc ccccctaacg ttactggccg aagccgcttg
     4861 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     4921 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     4981 cctctcgcca aaggaatgca gggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     5041 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     5101 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     5161 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     5221 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     5281 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     5341 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     5401 ctgttacatt gcacaagata aaaatatatc atcacgaaca gtaaaactgt ctgcttacat
     5461 aaacagtaat acaaggggtg ttatgggatc ggccattgaa caagatggat tgcacgcagg
     5521 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5581 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5641 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5701 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5761 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5821 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5881 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5941 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6001 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6061 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6121 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6181 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6241 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtct cccgcctccc
     6301 taccatccct gaggctcgct ggaaggcact gctcagagcc tgaagggctg acagcagaaa
     6361 agaggccctg gccagaggca gggtggccca tcggagagag cagtgacaca cccattgccc
     6421 gggggctgag tctcttcctc aacctcccac cagactctgc agaggcagcc cactgcccac
     6481 caccagccca aagaacctgc aggaaccttc cgcccgctga cctgcttgct ccagggtaac
     6541 tgtggaccct gtcctcgccc tgcgcacggt accctatgtc tggacacccg gccccagctc
     6601 cagccccagc ccaggtgggc cgcccctggc ggggtcgctt accaacggac acccacccca
     6661 gatgcatggg ccagagggcc ggcccccggc atagatgtgc acatcggttt tccagtgtga
     6721 acaaaagatt acgaaaccta gaaactgttg gttccgtgta agtagttgac tacgtgtttt
     6781 agaactgtgc tgaagacatc tgtaagacta ttttgtgggg gaaaaaagta gtttccttta
     6841 aggtaaaaag cattttatat gatccttagc acatttttaa gttttatctt aagggagacg
     6901 cgcacaaaag cggctgccaa accgtttcgt catcctcaca gcaaggaccg gacgcttgct
     6961 agccaccccg gagcactgct ctccttttaa tcatgtattc atctatttta aattgccggc
     7021 gacgactttt gtctatttat gaagaaacct tgagaacgaa gttacagctt atcctaccgt
     7081 gtgtgtggtt ttggggtttc gtttgggttt gggttcttga cgtcgtttgc agctgtttcc
     7141 tggccctggc gagtgtctgt cttggtgccc agtgcttctc tcaaatctct ttataataaa
     7201 acttctgaaa agctgaaaac aagtctgtgt ctccttgccc ttggtgagcc agggttgaac
     7261 cccctaaagt ggcatagagc ccctcgggcc agacggacac gggtcaaagt cttgcctgac
     7321 ctttaccagc ccatgtgggt ttgtgtgtgg gtgatggga
//
