LOCUS       sequence-229658-        6927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF193 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6927)
  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: pZNF193-donor
FEATURES             Location/Qualifiers
     source          1..6927
                     /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             3652..3672
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3685..3729
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3769..3792
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3820..3822,3823..3825,3826..4539)
                     /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(3865..3886)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4126..4145)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4577..5150,5151..5153,5154..5162)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4743..4760)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4970..4989
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5244..6047
                     /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(5307..5326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5917..5936
                     /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 aactacttgg gagactgagg caggagaatc
     2701 gcttgaacct gggaaatgga ggctgcagtg agctgagatc acaccattgc actccagcct
     2761 gggctacaga gtgacactct ggctcaaaaa acaaaacaaa aaacaacgac tcaggccaag
     2821 tctacatggg gatggactga cctgcattat gccaaagagg gcaaacgttc ctccctacgt
     2881 aggggaaagt gttaccacac tagcccttct gtttctagcc cacttccttt tctatgatat
     2941 atttttatag acatagtcac cttcccatag gctcagggaa caagtgacat ttacctagcc
     3001 tttttctttg tttcagatga agtaaccaag actgaggaca gagagttggt gctaaggaaa
     3061 gactgtccta agatagtgga accacatggg aaaatgttta atgagcagac ctgggaggta
     3121 tcacagcagg atccctcaca tggagaagtt ggtgaacata aggataggat agagaggcag
     3181 tggggaaacc tcttaggaga ggggcaacac aaatgtgatg aatgtgggaa gagctttact
     3241 cagagctcag gtctcattcg acatcaaaga attcatactg gagaaagacc ttatgaatgt
     3301 aatgaatgtg ggaaagcctt cagtcgaagt tctggtcttt ttaatcaccg aggaatccac
     3361 aatatacaga aacggtacca ctgcaaggag tgtgggaagg tcttcagtca gagtgcgggt
     3421 cttatccagc atcagagaat ccacaaagga gaaaagccgt atcagtgcag ccagtgcagt
     3481 aagagctaca gtcggcgttc atttctcatt gaacatcaga gaagccacac aggggagcga
     3541 cctcaccagt gcattgaatg tgggaaaagc tttaatcgac actgcaacct cattcgccat
     3601 cagaagatcc acacagtggc tgagctggtc gggattccaa ctactgcaag cgagaatctt
     3661 tattttcagg gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac
     3721 atggacagcg gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg
     3781 ttccaggggc ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa aggcgaggag
     3841 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3901 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3961 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     4021 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4081 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4141 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4201 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4261 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4321 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4381 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4441 ctgagcaaag accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc
     4501 gccgggatca ctctcggcat ggacgagctg tacaagtaat gatctagagt cgagttaatt
     4561 aagaattccg cccccccccc tctccctccc cccccctaac gttactggcc gaagccgctt
     4621 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     4681 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     4741 ccctctcgcc aaaggaatgc agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     4801 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     4861 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     4921 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     4981 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     5041 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     5101 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
     5161 cctgttacat tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca
     5221 taaacagtaa tacaaggggt gttatgggat cggccattga acaagatgga ttgcacgcag
     5281 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5341 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5401 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5461 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5521 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5581 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5641 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5701 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5761 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5821 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5881 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5941 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6001 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagtc tagggcttgg
     6061 ctatgagcaa gttttccaga tcaccaccca agttgtgtgg ggcaggttga gactagaaaa
     6121 tgcctctttc ttcctttctc catgaaatgt gtttgaaaca aatcctgact taaggcccag
     6181 ggacttcctt aaaggaaagt tgggtgtttg aagctactgt tttctctttt gttcatttta
     6241 cctctttctt actcttacta gctgtgtccc tcttatttat aatttattta tttttttgag
     6301 atggctgcta aacccttcta ataatataat aaatggcact gccacagcca acatgcctga
     6361 aagatttggt atctgtgttt aactacaaag gataacagag ttctactgat ggtttataac
     6421 aattgggtct ttggtgttcc ctgtgaccag ggtctacact ataaacacct tttctacctt
     6481 actgcttctc tctgacacct acgtgactct cccttggaaa gtgaccagca ttggctgtgt
     6541 gtttcagtgt ctgcagtgtg tgtccgctta cttaaccata gttggtcctc acaacagcca
     6601 cacagggaaa tgagcgttat cattatcgca ttttacagat gaggactcca aggcttagaa
     6661 aatccaagca tcagagccaa ggctgaaacc ctgaactttt atttttagta atgctttaat
     6721 tttggaataa tatttgactt acagaaatgt tgcagagatg gagttctcat attcccttca
     6781 cctagattct cctaatgtta acttgtaaca tcacaccttt ttagttaaac cttggctttt
     6841 aactccaaat ctcaatttct ttatgatgtc aagctttgta ctgataccag atttctacat
     6901 tttagtcacc tggaactccc ttagagt
//
