LOCUS       sequence-229749-        7278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF462 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7278)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7278)
  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: pZNF462-donor
FEATURES             Location/Qualifiers
     source          1..7278
                     /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             3872..3892
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3905..3949
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3989..4012
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4040..4042,4043..4045,4046..4759)
                     /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(4085..4106)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4346..4365)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4797..5370,5371..5373,5374..5382)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4963..4980)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5190..5209
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5464..6267
                     /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(5527..5546)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6137..6156
                     /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 tcggtacccg gtcacccagc cgcaagtcaa atagggaaaa
     2701 aaatcgtctt tatgaatgat cagtcctgct caagcctgtg tgcagaagta tagtgttttc
     2761 tccagccaat atccctttag tcttcattag cgtgacattc tataatagaa cgtgaatcga
     2821 ggtcaaaaga tctggcttct gttcctgcct ctctgactga tcattccctg agtggccctg
     2881 aacattccca tgtactgcct cggggacaga gccagcttct gcctgcctcc caggtaggtg
     2941 agagcatact tgtaaaatac ttatgagatc cctgaacata aaatcagccc ctcaacaagg
     3001 cacgttagtc caccagatgc agagagaacc taggatgtgg tcttttcagg aggaaacatg
     3061 gcttgtgtgg tgataggaag cctgaagctt ggtccctgaa gcctcaaggg tctagacctc
     3121 agagaatata accaagcttc ccgagacatg ggttccatct tcaggttttt gtttttagtt
     3181 tgttttcgtg atttgtttcg ggacctgaca aataaatctt tagtacctga gttactcttt
     3241 tagacttcat aaatataacc aatctccaag cttctgggtt cagccgatga ctctgagcat
     3301 aagacaattc ttgctgttac cacttatgac ccagtagaga aagtgacata aaatgattat
     3361 gcttatctag aaagaaaaca tgggataatc aatatatagt ataataatgg atacttctgt
     3421 aagttcaaat ctttaagata cagaaatata tgacttaagc aaaacatagc agggatgcat
     3481 cacttggtat tttgtcatac acccatggca tttgttcaaa ggaaacccca aggctttttg
     3541 aggatcagga aaataaagac actcgagggt ttttattatt tttttgttta aaaagagaaa
     3601 tatatatact atactcatct gtctcttcga taaatgtttt tccagggcat tgaatgacac
     3661 caagcaggtg agcagagaag aaatccaccc aaaagagatc atggagaaca gtgttaaaat
     3721 gccctccata gaggaaaagg aagatgacga ggccattggg atagactttt ccctaaagaa
     3781 tgaaacagta gccatctgtg tagtaactgc cgacaaatct ctcctggaga atgcagaggc
     3841 caaaaaagaa gggattccaa ctactgcaag cgagaatctt tattttcagg gcgccgccaa
     3901 attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg gaggtggagg
     3961 ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc ccctgtcgtc
     4021 gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg gggtggtgcc
     4081 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4141 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4201 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4441 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4501 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4621 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4681 gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca ctctcggcat
     4741 ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg cccccccccc
     4801 tctccctccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4861 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     4921 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     4981 agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     5041 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     5101 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     5161 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     5221 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     5281 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     5341 tggttttcct ttgaaaaaca cgatgataat atggccacaa cctgttacat tgcacaagat
     5401 aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa tacaaggggt
     5461 gttatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5521 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5581 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5641 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5701 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5761 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5821 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5881 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5941 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6001 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6061 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6121 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6181 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6241 tatcgccttc ttgacgagtt cttctgagtc gcgtttggtg aaattcttaa tcaaacctta
     6301 cttgaacagt gatgaaaaag tgggagggct ggcttgggct gagaagggag ggacagaaaa
     6361 gagaagacag aacaaagctg ctttttagga ctgaacaatc tattttcaaa gcactggtac
     6421 ctgtgtgagt gagtatgtaa attaaagtta tttaaatggt tggaatatgt ggctcctttt
     6481 ccatcactac atcttttctt ccggatcttc atcatggaag tttcatttgt tgcggaatat
     6541 ggaagcacct cccaatggta cggtgcaccc tgtggtggtc ttggacagta tgtggaaaca
     6601 gaagctccat gacggtagaa gacttctcat tggggagcaa ctttttgacg cacaactttt
     6661 ggtgcgtttt tctagtttta ataccttaag ctttttcaag acctaactgc agccgctttg
     6721 ggaaaaaaaa acaaaaaaca aaaaacagaa aacaaaaaaa aaaaaactgc tttgcataaa
     6781 acagtcacct gtttcctagt acctcaaact taaccaaggg aactctctgc atttgtcagt
     6841 actacctgtc gtcgttgtgg ttaaatgtag agagaactgc tgggcaagat ggtgaatgga
     6901 gaaaaaaaaa gagttttaaa gaaaggaaca caaattgtgc ttaaaatccc catgtgggtt
     6961 ttatttctta aaatactgtg atttttttaa ttattttagt aaaaaacaaa acaaaaaaaa
     7021 gaaacagact ttaattatta gataactgtt tgtgaattgt catcctttat tccaggtaag
     7081 ctgtttatta gtgttcaact ataatttaga atttggtaga ttcatgtgag ctaattttaa
     7141 ggtgattgtg gagttttgtt tgccacatgt ttattttcta tcaatttgag tgttacaaac
     7201 acagcacaat tcagtttttc atatgaattg tttttcgtgt gtgtgttgtt attgttgttt
     7261 taatttcggg gcaaggga
//
