LOCUS       sequence-229838-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF250 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7227)
  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: pZNF250-donor
FEATURES             Location/Qualifiers
     source          1..7227
                     /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             3886..3906
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3919..3963
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4003..4026
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4054..4056,4057..4059,4060..4773)
                     /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(4099..4120)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4360..4379)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4811..5384,5385..5387,5388..5396)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4977..4994)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5204..5223
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5478..6281
                     /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(5541..5560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6151..6170
                     /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 caaagcttct gtctgagtcc aaactctgtt
     2701 gaccaccgtg aagttcaggt cttaagccaa agcatgccac tcactccgca ccaggcagtg
     2761 cctagtggag agaggcccta catgtgtgtt gagtgtggga agtgctttgg ccggagttcc
     2821 cacctccttc agcatcagcg tatccacact ggagagaagc cctatgtgtg cagtgtatgt
     2881 gggaaggcct tcagccagag ctcagtcctt agtaaacaca ggagaattca cacaggtgag
     2941 aagccctatg agtgtaatga gtgtggaaaa gcctttagag tgagctcaga tcttgctcag
     3001 catcacaaga tacatacagg agagaagcct cacgaatgtc ttgagtgtcg gaaagccttc
     3061 actcaactct cacatctcat tcagcaccag cggatccaca cgggagaaag gccatatgtg
     3121 tgtccgttgt gtgggaaagc cttcaaccat agcactgttc tgcggagcca ccagagggta
     3181 cacactgggg agaagcctca caggtgcaat gagtgtggga aaaccttcag tgtgaagagg
     3241 acactgctgc agcaccagag gatccacacc ggggagaagc cctacacgtg cagcgagtgt
     3301 gggaaggcct tcagcgaccg ctcagtcctc attcagcacc acaacgtgca caccggggag
     3361 aagccctatg agtgcagtga gtgtgggaag accttcagcc accgctccac actgatgaat
     3421 cacgagcgga tccacaccga ggaaaagccc tatgcatgct acgaatgtgg gaaggccttc
     3481 gttcagcact cacacctgat ccagcaccag agagtccaca ctggggagaa gccctatgtg
     3541 tgtggtgaat gtgggcacgc cttcagtgca cgccggtctc tgatccagca tgagagaatc
     3601 cacacaggtg aaaagccctt ccagtgcaca gaatgtggca aagccttcag cctgaaagca
     3661 actctgattg tgcacctgag gacccacacg ggcgagaagc catatgagtg caatagctgc
     3721 gggaaggcct tcagccagta ctcagtgctc atccagcacc agcggatcca cacaggcgag
     3781 aagccctatg agtgcgggga gtgtgggcgt gccttcaacc agcatggcca cctaatccag
     3841 caccagaaag tgcacagaaa gttggggatt ccaactactg caagcgagaa tctttatttt
     3901 cagggcgccg ccaaattcaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac
     3961 agcggaggtg gaggttcgag cggtccctcg ggttcgtcga gcctggaagt tctgttccag
     4021 gggcccctgt cgtcgagcgg tccctcgggt tcgatggtga gcaaaggcga ggagctgttc
     4081 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4141 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4201 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     4261 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     4321 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     4381 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     4441 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     4501 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     4561 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     4621 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     4681 aaagacccca acgagaagcg cgatcacatg gtccttaagg agttcgtgac cgccgccggg
     4741 atcactctcg gcatggacga gctgtacaag taatgatcta gagtcgagtt aattaagaat
     4801 tccgcccccc cccctctccc tccccccccc taacgttact ggccgaagcc gcttggaata
     4861 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     4921 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     4981 cgccaaagga atgcagggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     5041 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     5101 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     5161 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     5221 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     5281 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     5341 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc acaacctgtt
     5401 acattgcaca agataaaaat atatcatcac gaacagtaaa actgtctgct tacataaaca
     5461 gtaatacaag gggtgttatg ggatcggcca ttgaacaaga tggattgcac gcaggttctc
     5521 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5581 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5641 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5701 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5761 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5821 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5881 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5941 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6001 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6061 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6121 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6181 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6241 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agtccccatg gctgacacaa
     6301 gaatccattc tcacagaaac tgcatgtgga accacaagca gccttcagcc caagagaagt
     6361 ctctgttaac tctataggaa gcttttcttt ggcgattcag tgtcacaaaa taactccaga
     6421 aagaagcact tagcgtgctg ttcctgtgga aaaacttcag agactacctg ttttattttc
     6481 ctcaacatct tgaagttatg ttggagagta atcatacaat tgtagagaat tttggtaaaa
     6541 aacagccata attctttaac attagtttat ttgaactaag ggaatttaag gcataagaac
     6601 cattatccca ataaaatctt acattccaaa taaagttctt tttctaagaa cattacatgc
     6661 ccttcctaaa tatcaattaa ccatactaat tattgcactt aaaatttgaa agtcggacta
     6721 ttttcagtat tctcttaaaa gactaaagta ttgtatggat aaagtgatat aaaaagatat
     6781 tttcatcaag catcatgtaa aatggttgga aatcctaagt gtgtggattt ccatcctaag
     6841 tgtgtggagt cctgttttgt atgaaacaag gaaaaagctt atatattagt gagaaattac
     6901 ataaatttaa aaaacttttt gtatgtgtgt gtggagatag tcttgctctg ttgcccaagc
     6961 tgcagtgcag tggcatgatc ttggctcact gcagtctccg cccccacccc aggttcaagc
     7021 ttttcttgtg cctcagtctc ccaagtaact aggattatag atatgcgcca ccacacctgg
     7081 ctaatttttg tatttttagt agagacggag tttcaccgtg ttggccaggc tggtctcgaa
     7141 ctcctggcct caagtgatcc tcccaccttt gcttccgaaa gtgttgggat tacaggcgtg
     7201 aggcaccatg cctggcctag aaaaccc
//
