LOCUS       sequence-211740-        7068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF695 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7068)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7068)
  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: pZNF695-donor
FEATURES             Location/Qualifiers
     source          1..7068
                     /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             3623..3643
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3656..3700
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3740..3763
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3791..3793,3794..3796,3797..4510)
                     /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(3836..3857)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4097..4116)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4548..5121,5122..5124,5125..5133)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4714..4731)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4941..4960
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5215..6018
                     /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(5278..5297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5888..5907
                     /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 tcggtaccct ccatattgga gagaacccgt accaatgtaa
     2701 aaaatgtggc aaagccttta atgagtgctc atgctttact gactgtaaga gaattcatgt
     2761 tggagagaaa cattgcaaat gtgaagaatg taataacatt tttaagtctt gctcaagtct
     2821 tgctgttgtt gagaaaaatc atactgaaaa gaaaacctac agatgtgaag aatgtggcaa
     2881 agcttttaac ctgtgctcag ttcttactaa acataagaaa attcatactg gagagaaacc
     2941 atacaaatgt gaagaatgtg gcaaatcctt taagttgttc ccatacctta ctcaacacaa
     3001 gagaattcat agtagagaga aaccctacaa gtgtgaagaa tgtggcaaag tctttaaatt
     3061 gttgtcatac cttactcaac atagaagaat tcatactgga gagaaaacct tccgatgtga
     3121 agaatgtgga aaagccttta accagagctc acatctgact gaacatagga gaattcatac
     3181 tggtgagaaa ccatacaaat gtgaggaatg tggcaaagct tttacctggt tctcatacct
     3241 tattcagcat aagagaattc atactgggca gaaaccctac aaatgtgagg aatgtggcaa
     3301 agcttttacc tggttttcat accttactca acataagaga attcatactg gagagaaacc
     3361 ctacaaatgt gatgaatgtg gcaaagcttt taactggttt tcatatctta ctaatcataa
     3421 gagaattcat actggagaga aaccctacaa atgtgaagaa tgtggcaaag cctttggcca
     3481 gagctcacac ctttctaaac ataagacaat tcataccaga gagaaaccat acaagtgtga
     3541 ggaatgtggc aaagccttta accactctgc acaacttgct gtacatgaga aaactcatac
     3601 cgggattcca actactgcaa gcgagaatct ttattttcag ggcgccgcca aattcaaaga
     3661 aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag gttcgagcgg
     3721 tccctcgggt tcgtcgagcc tggaagttct gttccagggg cccctgtcgt cgagcggtcc
     3781 ctcgggttcg atggtgagca aaggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3841 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3901 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3961 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4021 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4081 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4141 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4201 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     4261 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     4321 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4381 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     4441 tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4501 gtacaagtaa tgatctagag tcgagttaat taagaattcc gccccccccc ctctccctcc
     4561 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     4621 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     4681 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg cagggtctgt
     4741 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     4801 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     4861 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     4921 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     4981 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     5041 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     5101 tttgaaaaac acgatgataa tatggccaca acctgttaca ttgcacaaga taaaaatata
     5161 tcatcacgaa cagtaaaact gtctgcttac ataaacagta atacaagggg tgttatggga
     5221 tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5281 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5341 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5401 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5461 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5521 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5581 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5641 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5701 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5761 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5821 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5881 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5941 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6001 cttgacgagt tcttctgagt ctgagaaaaa ccctacaaat ctaaacaata tggcatagtc
     6061 tttaatacct attcacaact tcacagcaga atatttttac tgaataagag tgttacaaat
     6121 gtaatgactg tcaaaaggcc atttacagtc tatgagcctt tgagtgcact aaaatgttta
     6181 ggctacgaac aatacaaata gaccggttca acacctccac ttatatcaca gctcttactg
     6241 tacacagaag aatttatact ggagggaaac cctccagttg ctcaaactgt attcaatttc
     6301 aaagactttg tattggagag aaaccctaca aatgtaataa atgcagaaac aacatttgtt
     6361 caaaaaatat acctcagaaa acaccagagt gttcacacta aaaactgttt tacagatgca
     6421 gtaaatgtga aaaaatgtct aatcaaaaat tacatcaaaa cacatccaag aattcatagt
     6481 aaaaagcact aagtcactga cactttcaga cattactgta aatctgagtg ttggttatag
     6541 agaataattc aaagttaagt taaagtaagt aggagattca ccttttgggg aagttataat
     6601 tacatttcaa gtataccttt tggtgccagg cacggtggct ctttcctata gttgctgcac
     6661 ttttggatgc cgaggtcggg ggattgcttg agcccaggag tttgggacca ggctgggcaa
     6721 catggcaaaa cctcatctct acaaaaagta aaataaaagc caggcttggt ggcacatgcc
     6781 tgttgtccca gctacttgga aggctcaggt gggaggattg cttgagcctg gaggttgagg
     6841 atgcagtgag ctgctatctg gcaacttcac tcctgcctgg gcaacagagc aagacccttt
     6901 ctcaataata ataataacaa cataataata aagtatactc ggtgcactga aagagtttta
     6961 gcttttttga aaatcacata tttatgtaat tcaagtctta aatcacttga taccatgcct
     7021 tcatttctag tgtttatgtg aaggcatgag gcctactgtt gctacatg
//
