LOCUS       sequence-228576-        7194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF354C with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7194)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7194)
  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: pZNF354C-donor
FEATURES             Location/Qualifiers
     source          1..7194
                     /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             3711..3731
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3744..3788
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3828..3851
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3879..3881,3882..3884,3885..4598)
                     /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(3924..3945)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4185..4204)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4636..5209,5210..5212,5213..5221)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4802..4819)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5029..5048
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5303..6106
                     /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(5366..5385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5976..5995
                     /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 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 tcggtacccc ctcacatctg taatgaatgt gggaagagct
     2701 tcaagcagaa tctgcatctt attgaacatc agagaattca tacaggtgag aaaccctaca
     2761 aatgtaatga gtgtgaaaaa accttcagcc acagatcatc ccttctttct catcagagaa
     2821 ttcatactgg agagaaacct tacaagtgta atgaatgtga gaaggcattt agcaacagtt
     2881 caacccttat caaacatctg agagtgcata ctggagagaa accgtatcga tgtagggaat
     2941 gtggtaaagc ctttagccag tgttcaaccc tcactgtaca tcagagaatt catactggag
     3001 agaaactcta taaatgcggc gaatgtgaga aggccttcaa ctgtagagca aaacttcaca
     3061 ggcatcaaag aatccataca ggtgagaaac cctataaatg tagtgagtgt gggaagggat
     3121 acagccagtt tacatctcta gctgaacatc agaggtttca tactggagaa caactgtata
     3181 catgcttgga atgtgggaga accttcacac gtattgtaac ccttatcgaa catcagcgaa
     3241 ttcacactgg acaaaaacct tatcagtgca acgaatgtga gaaagccttc aaccagtatt
     3301 catcctttaa tgaacatcgg aaaattcata ctggggaaaa actttataca tgtgaggaat
     3361 gtgggaaagc ctttggttgc aaatctaacc tttataggca tcagagaatt catactggag
     3421 agaaaccgta tcagtgtaat cagtgtggaa aggccttcag ccagtattca tttttaaccg
     3481 aacatgagag gatccacact ggagagaaac tgtataaatg tatggaatgt gggaaagcct
     3541 acagttacag atcaaacctt tgtagacaca aaaaagttca cacgaaagag aaactctata
     3601 agtggaagga atatgggaaa cctttcatct gcagctcctc acttacccag tatcagagat
     3661 tttttaaagg agataaagcc tatgaggtcg ggattccaac tactgcaagc gagaatcttt
     3721 attttcaggg cgccgccaaa ttcaaagaaa ccgctgctgc taaattcgaa cgccagcaca
     3781 tggacagcgg aggtggaggt tcgagcggtc cctcgggttc gtcgagcctg gaagttctgt
     3841 tccaggggcc cctgtcgtcg agcggtccct cgggttcgat ggtgagcaaa ggcgaggagc
     3901 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3961 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4021 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4081 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4141 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     4201 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4261 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     4321 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     4381 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     4441 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     4501 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct taaggagttc gtgaccgccg
     4561 ccgggatcac tctcggcatg gacgagctgt acaagtaatg atctagagtc gagttaatta
     4621 agaattccgc ccccccccct ctccctcccc ccccctaacg ttactggccg aagccgcttg
     4681 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     4741 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     4801 cctctcgcca aaggaatgca gggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     4861 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     4921 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     4981 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     5041 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     5101 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     5161 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     5221 ctgttacatt gcacaagata aaaatatatc atcacgaaca gtaaaactgt ctgcttacat
     5281 aaacagtaat acaaggggtg ttatgggatc ggccattgaa caagatggat tgcacgcagg
     5341 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5401 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5461 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5521 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5581 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5641 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5701 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5761 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5821 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5881 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5941 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6001 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6061 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtct agttcatctc
     6121 tcaaataatc caagacttct cactggggaa taagggaata ataaataggg tacaaactcc
     6181 taatagattt gtctttttta cttctcctga aggaaatatg ttagttgcca ctaagtcatg
     6241 ataaaattga tcagtgagac tatgaagagc actgacttgt taaattttaa aagaaccata
     6301 aattctaagg tatctaaaaa cctatgagta tttaattcat agaaaaaatg taaaaggtct
     6361 ttttaaaaat catgaaaaat agttgaatat acattttgtt tctctcataa gaccatattc
     6421 cctttaaaag agtaagcttc aatatgtgaa ttttctttta aaaacagtca ctgagttaat
     6481 aatgtaaata agtgtgtggc cttctttaaa atagctggct aacataggag gcacttcttt
     6541 tcataaagag aagctaaaca taaaaaggaa ttttaaattt aactcttcac atggaaataa
     6601 taaagctctt tatatgagct gtcccaccag caacttatat atgtaaacat acatatatac
     6661 acatatgcat gtgtgtgtgt aaacataaaa gtcctttatt attaaaaaat gtagttttat
     6721 tgatttagaa tttagttctt aaaatcttgc ccaaaggtag actctctttc ttttactttc
     6781 ttccattatt tcttaaattg atgtggtttt atataactga acagagtact attacagtac
     6841 cgtagaaggc ggattaactg aagcagctgg aaagactgtc aagcacatac tttgttatct
     6901 gttgcagaag ggatgtattt ggcagaatct caggctgcat gtctggtagc tcattagagc
     6961 caggagagat tttagtttaa ttatagcctt tcattgtact aacgaaggat ttgaacttag
     7021 tgaacagagt caagactaga atctaggtct ttggttctgg tgttttgtga tcattatagg
     7081 ctcgtttcca acctgaggtc ccaagagtac tatttgtttt gtagcctggt tagacatagt
     7141 tgcagtaatt taaagttact gtgttatctt taagggcttc ctctcacctt aatc
//
