LOCUS       sequence-211768-        7137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor TFE3 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7137)
  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: pTFE3-donor
FEATURES             Location/Qualifiers
     source          1..7137
                     /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             3882..3902
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3915..3959
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3999..4022
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4050..4052,4053..4055,4056..4769)
                     /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(4095..4116)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4356..4375)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4807..5380,5381..5383,5384..5392)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4973..4990)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5200..5219
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5474..6277
                     /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(5537..5556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6147..6166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gacctgcagg 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 tagaaggaca 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 ggtggaggca gagagagagc aggccagggt
     2701 ttcccttcct ggcatacttg ggggatccct gggcttccca gatcacacga tgatgatagt
     2761 gataatgata aataataatc ataatattaa caaggataat aatattgcta atgtacatag
     2821 catttatctt gtgctctgta ctgtgacaag agctttatat ccactaactt attcaatcct
     2881 cagaattcta tgagataggg gccattattt tatttattta tttatttatt tatttttgag
     2941 atggagtttc gctcgttgcc caggctggag tgcaatggcg caatctcggc tcaccgcaac
     3001 ctccgcctcc cgggttcaag caattctcct gtctcagcct cctgagtagc taggattata
     3061 ggcatgcacc accacaccca gctaattttt gtatttttag tagagatggg gtttctccat
     3121 gttggtcaag ctggtctcga actcctgacc tcaggtgatc cacctgcctc ggcctcccaa
     3181 agtgattaca ggcgtgagcc accgcgccca gcctaggggc cattattata tacccacttt
     3241 acagtttgcg actctgaagc ttcagagatg ttaagcacct tgcctaaagt tttggaacta
     3301 gtgagtagca gaagctgaat tccaacccag ggaacctacc cttttcaacc tttatactcc
     3361 tctgcctctg agggactctg aaccctactt atcctgcacc caacccctat tttttacagg
     3421 aactagaact gcaggcccag atccatggcc tgccagtacc tcccactcca gggctgcttt
     3481 ccttggccac gacttcggct tctgacagcc tcaagccaga gcagctggac attgaggagg
     3541 agggcaggcc aggcgcagca acgttccatg tagggggggg acctgcccag aatgctcccc
     3601 atcagcagcc ccctgcaccg ccctcagatg cccttctgga cctgcacttt cccagcgacc
     3661 acctggggga cctgggagac cccttccacc tggggctgga ggacattctg atggaggagg
     3721 aggagggggt ggtgggagga ctgtcggggg gtgccctgtc cccactgcgg gctgcctccg
     3781 atcccctgct ctcttcagtg tcccctgctg tctccaaggc cagcagccgc cgcagcagct
     3841 tcagcatgga agaggagtcc gggattccaa ctactgcaag cgagaatctt tattttcagg
     3901 gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg
     3961 gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc
     4021 ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg
     4081 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4141 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4201 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4261 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4321 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4381 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4441 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4501 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4561 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4621 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4681 accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca
     4741 ctctcggcat ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg
     4801 cccccccccc tctccctccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     4861 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     4921 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     4981 aaaggaatgc agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     5041 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     5101 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     5161 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5221 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5281 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5341 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa cctgttacat
     5401 tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa
     5461 tacaaggggt gttatgggat cggccattga acaagatgga ttgcacgcag gttctccggc
     5521 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5581 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5641 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5701 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5761 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5821 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5881 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5941 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6001 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6061 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6121 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6181 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6241 catcgccttc tatcgccttc ttgacgagtt cttctgagtc tcaggcctca cccctcccct
     6301 gggactttcc cacccaggaa aggaggacca gtcaggatga ggccccgcct tttcccccac
     6361 cctcccatga gactgccctg cccaggtatc ctgggggaag aggagatgtg atcaggcccc
     6421 acccctgtaa tcaggcaagg aggaggagtc agatgaggcc ctgcaccttc cccaaaggaa
     6481 ccgcccagtg caggtatttc agaaggagaa ggctggagaa ggacatgaga tcagggcctg
     6541 ccccctgggg atcacagcct cacccctgcc cctgtgggac tcatccttgc ccaggtgagg
     6601 gaaggagaca ggatgaggtc tcgaccctgt cccctaggga ctgtcctagc caggtctcct
     6661 gggaaaggga gatgtcagga tgttgctcca tcctttgtct tggaaccacc agtctagtcc
     6721 gtcctggcac agaagaggag tcaagtaatg gaggtcccag ccctgggggt ttaagctctg
     6781 ccccttcccc atgaaccctg ccctgctctg cccaggcaag gaacagaagt gaggatgaga
     6841 cccagcccct tcccctggga actctcctgg ccttctagga atggaggagc caggccccac
     6901 cccttcccta taggaacagc ccagcacagg tatttcaggt gtgaaagaat cagtaggacc
     6961 aggccaccgc tagtgcttgt ggagatcaca gccccaccct tgtccctcag caacatccca
     7021 tctaagcatt ccacactgca gggaggagtg gtacttaagc tcccctgcct taacctggga
     7081 ccaacctgac ctaacctagg agggctctga gccaaccttg ctcttgggga aggggac
//
