LOCUS       sequence-241245-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor E2F3 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7065)
  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: pE2F3-donor
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1100..1120
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1133..1177
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1217..1240
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1268..1270,1271..1273,1274..1987)
                     /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(1313..1334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1574..1593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(2025..2598,2599..2601,2602..2610)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2191..2208)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2418..2437
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2692..3495
                     /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
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLFDEFF"
     primer_bind     complement(2755..2774)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3365..3384
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4466..4482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4466..4482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4479..4501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4490..4506
                     /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(4514..4520,4521..4538,4539..4544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4559..4580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4697..4714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4868..5456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4948..4967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5627..6418,6419..6487))
                     /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     6250..6269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6488..6592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6660..6678
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6716..6738)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6838..6857
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7051..7065,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7065,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacccccag gagttcgagg ttagagtgag
       61 ctatgcttgc ctgtgaatag cccctacact ccagcgtaga caacatagca agacgccgtc
      121 tcttagaaaa aaaagttctt ttttcagttg gtgagtttct actaggaagc caacttttca
      181 gtatgtatat attgttttgt tttgattttc ttaaaacaat gactagatag cagagagaag
      241 tttttgaact tgttatcctc tcatcattga agttatttgc attcttgttt attgacagta
      301 gttatttgaa aaatgagtga gtagactagg gccaaagaaa gatggccagg catggtggct
      361 cacacctgta atttcagcac tttgggaggc caaggaagac agatcacttg agcccaggag
      421 ttcgagacca gactgggcaa catggcaaaa ccctgtctct acaaaaaaag attaaccagg
      481 catggtggct cacacctgga gtcccagcta cttgggaggc tgaggtggga gggtcccttg
      541 aacccagaag ttcaagactg cagtgagtca tgatcacacc actgcactcc agccaagaga
      601 ataagaccct gtctcaaaaa ttaaaagggg gacgggggtg gcaaggaact tttttgtacc
      661 tgttagcata aaaggtgatc tgcatcataa agtcgtctca ttgttattat ttgcggaagg
      721 tacatgcttt tttctaacag caacatatac attcttccag aaaaattcat ttgatttttc
      781 taacttattt tttgtttcca tcaatgtttt cttttcagac ttggcttcaa ccaactcagg
      841 acatagcgat tgctcagttt ctatgggaaa cctttctcct ctggcctccc cagccaacct
      901 cttacagcag actgaggacc aaattccttc caacctagaa ggaccgtttg tgaacttact
      961 gcctcccctg ctgcaagagg actatctcct gagcctcggg gaggaggaag gcatcagcga
     1021 tctcttcgat gcttacgatt tggaaaagct cccactggtg gaagacttca tgtgtagtgg
     1081 gattccaact actgcaagcg agaatcttta ttttcagggc gccgccaaat tcaaagaaac
     1141 cgctgctgct aaattcgaac gccagcacat ggacagcgga ggtggaggtt cgagcggtcc
     1201 ctcgggttcg tcgagcctgg aagttctgtt ccaggggccc ctgtcgtcga gcggtccctc
     1261 gggttcgatg gtgagcaaag gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1321 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1381 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1441 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1501 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1561 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1621 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1681 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1741 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1801 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1861 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1921 catggtcctt aaggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1981 caagtaatga tctagagtcg agttaattaa gaattccgcc cccccccctc tccctccccc
     2041 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2101 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2161 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcag ggtctgttga
     2221 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2281 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2341 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2401 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2461 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2521 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2581 gaaaaacacg atgataatat ggccacaacc tgttacattg cacaagataa aaatatatca
     2641 tcacgaacag taaaactgtc tgcttacata aacagtaata caaggggtgt tatgggatcg
     2701 gccattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2761 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2821 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2881 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2941 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3001 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3061 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3121 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3181 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3241 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3301 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3361 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3421 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttttt
     3481 gacgagttct tctgagtctg attatgcttc gtgtgaactc tccttaaaaa ccgatattta
     3541 tttatcatgg aaccagaaca tctgtcatgc agtgttgtcc cttcctacct tcttcctcca
     3601 agagagtgtc atgaagtaaa ctacaaactt cagaagaaag ctgacatttt aatgaatttt
     3661 ttaaaaaatt aataaacaaa ttgtctaaac gcacagttgc aggctccctt gggaaagccc
     3721 tgctttgctc caggctccaa gatctcctgg ctaagtcagc aagtgagaaa atgtgcaatc
     3781 aggtgtctct caccccgaat tgtccttcct ccttcctccc cggattggct tgctgtgcct
     3841 gacggatggg ctgtagaatg gggtctggcc acctggcctg ctgggaaaca gcaatcttcc
     3901 ttaatagcat ttcaagccgt gccttctccg cagaatgcat gtctttgagg tctgctaata
     3961 tggaatggaa ctgcagcaaa tgcaaacttg aagtcatgca aaagtatgaa atggatttct
     4021 tcagctcttc ttaggaatat ttaaattact gtcataattc agtttaagcc atgaactgtg
     4081 tgtcccagta ggaggtcaag aaaacctcca cagccttctg gatgaagaac ctgttttcaa
     4141 atatacttgt tgcagataca gaagactagt agagttctgc cactctaagc tgttgtggat
     4201 tttcctgtct ccatgaacca ctcccattcc cccgtcccca atgtgtttgt gagtttccag
     4261 ttgatttgta gcaaatgcct acttagttct ttgtggattg ttctagactt ttaatttttt
     4321 tagctgccat ttaagcattc ctgtggcacc catcaccatt tcaatttaat tgttttcttt
     4381 gaagcggttt ttgcaaattc atattactta agcagaggga gagaacctct acgacctgca
     4441 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4621 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4681 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4741 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4801 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4861 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4921 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4981 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5041 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5101 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5161 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5221 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5281 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5341 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5401 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5461 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5521 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5581 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5761 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6001 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6601 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6661 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6721 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6781 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6841 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6901 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     6961 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     7021 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt
//
