LOCUS       sequence-214769-        7348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor TCFL5 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7348)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7348)
  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: pTCFL5-donor
FEATURES             Location/Qualifiers
     source          1..7348
                     /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             3878..3898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3911..3955
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3995..4018
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4046..4048,4049..4051,4052..4765)
                     /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(4091..4112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4352..4371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4803..5376,5377..5379,5380..5388)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4969..4986)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5196..5215
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5470..6273
                     /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(5533..5552)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6143..6162
                     /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 tcggtacccc cccttccttt cttcctcacc gttcatttgt
     2701 tgaaaaggga atgatttgtc ccgtagacgt ttccacattc tgcccgtggt ggttggcgcc
     2761 tgtcatcggg ccgtcgcctg tgcccctctc ttcctctcag tctccttcgg gaggttgaga
     2821 ctgaccccac cattataatt gaggcctttt gtaaaagaca ggtggtgtga caggagcaag
     2881 ctcagcccac ccccaccctg ctggcagggt ctccccaagg acagagccat ctcagcgagc
     2941 agcacctcca ccttgctggc cttggaaccc tgagtgtgtg gcagtgcaga gggcaaggca
     3001 gtcgccctgg ggtgggggta gatgcccctg acttgttaat tgttgaaact ttttaaaaat
     3061 ttattttttt gagacggagt ttcactcttg ttgcccagac tggagtgcaa tggcatgatc
     3121 tcggctcact gcaacctcca cctcccaggc tcaagagatt gtcctgcctc agcctcccaa
     3181 gtagctgggg tcacaagatg cttgccacca cgcccagcta atttttgtat ttttagtaga
     3241 gatggggttt caccatgtta atcaggctgg tcctgaactc ctgacgtcag gtgatccacc
     3301 cacctcggcc tcccaaagtg ctgggattat aggcgtgagc cgccacgccc ggcctgaaac
     3361 gttttaatat atacactcta ccttctctca cacgctgggt ttacaggttt attagctagg
     3421 gcccattagc ggttgaaacg ttcatgtcat gtatatttta ccacaataaa atcaatacag
     3481 tgtggccaga cattctgagt attggaaaaa caaaagtaca tgtacagtgt aactgctatt
     3541 tgaaaaaaaa aaagttaaaa tctgtttatt atataaatcc ccttgtgagg taagacacaa
     3601 gtttgtgaat gacaccttaa aacagcgtat tctaaagtca tttacttagc tcttaatttg
     3661 aaccttttgc actttggaag acagcagttt tcatgtcagt taattcagta tgttaaccgt
     3721 cacttcttca ctgcaggaat ttgagagcgt attttgcggt aaaactggcc gaaggctaaa
     3781 gctgaccaga ccggactcct tggtgacctg tcctgcacag gggagtttac agagcagccc
     3841 ctcgatggag atcaagggga ttccaactac tgcaagcgag aatctttatt ttcagggcgc
     3901 cgccaaattc aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg acagcggagg
     3961 tggaggttcg agcggtccct cgggttcgtc gagcctggaa gttctgttcc aggggcccct
     4021 gtcgtcgagc ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt tcaccggggt
     4081 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4141 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     4201 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     4261 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4321 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4381 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4441 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4501 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     4561 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4621 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4681 caacgagaag cgcgatcaca tggtccttaa ggagttcgtg accgccgccg ggatcactct
     4741 cggcatggac gagctgtaca agtaatgatc tagagtcgag ttaattaaga attccgcccc
     4801 cccccctctc cctccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     4861 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     4921 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     4981 gaatgcaggg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     5041 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     5101 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     5161 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     5221 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     5281 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     5341 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacctg ttacattgca
     5401 caagataaaa atatatcatc acgaacagta aaactgtctg cttacataaa cagtaataca
     5461 aggggtgtta tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct
     5521 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5581 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5641 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5701 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5761 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5821 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5881 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5941 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6001 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6061 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6121 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6181 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6241 gccttctatc gccttcttga cgagttcttc tgagtctcgg actgaacagg aatcctcggg
     6301 gggtgaacag ccattccttc gtgacctgtg cacgccttct gcaaccctgg agctctgctc
     6361 ggctagtctg actcgaaaag ggcgtgactc aagctgacgg gactccagta gggactttga
     6421 gagcacattt tgtaaaaata tttatctaga cgcaaatgct tatccatgaa tgtcctctta
     6481 gaccatttgg ggatgaagcc atcttaataa ttagtaataa ttaattagta ataattagta
     6541 agcattttct caatgctctg attccatcat gttttcttaa catgataact taaaaaattg
     6601 acatcctttg tactttcttt aatcttaaaa agtacacggc tttttactta tttacctttt
     6661 aaatatgccc ctttagcaat tggaacaagt taaattgtta actaaaaaca gtttggaaat
     6721 tttatttcat tcgttatatc acaccccctt gtcatgactc tgagtcacgt gctgctgtat
     6781 tgcaacgtgc aggaccattt taaacctgtg tgctaaaaat tttccagata cttgctttaa
     6841 agctactttt gtccacaaat gaaatactgt cacagtagac gcttaaatgc cacgttttca
     6901 taccaagagt cattcattac ttcatgtgtc acaaactgtg gtgtttggaa ttgggttttt
     6961 caatgagtgg ctttacttat caatcacaac aggtaatagc aatagacgtt agtgcaatac
     7021 aaagtcaccc tcaataaata ctgttaattg gagatgtgag tttgtacaca aaacatcaga
     7081 ctagaccttt gtatgggaga gaatttactg tacattaaat tctttatttt ttgttactct
     7141 gtagccagtc attaatctga aggtttaata tatcatttta ttgggatgag atcatagtct
     7201 ttacacaaat gctatgtaaa caagttactg aatatttttc acctcgtgga gttgtacaca
     7261 accttttata tatacacacc ctaccttctc tcaaatgctg ggcttacagg tttattagct
     7321 agggcctttt gaggtatgct gtcaggcg
//
