LOCUS       sequence-229718-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZKSCAN4 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7126)
  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: pZKSCAN4-donor
FEATURES             Location/Qualifiers
     source          1..7126
                     /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     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             3777..3797
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3810..3854
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3894..3917
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3945..3947,3948..3950,3951..4664)
                     /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(3990..4011)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4251..4270)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4702..5275,5276..5278,5279..5287)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4868..4885)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5095..5114
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5369..6172
                     /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(5432..5451)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6042..6061
                     /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 cagccacgac gttgtaaaac
     2641 gacggccagt gaattcgagc tcggtacccc cctccactcc cagtttgtac ccttccaatg
     2701 gccccattct cattctcagg cctccttcct ccccagcacc cctgttctcc ctttttattc
     2761 ccattcttat tctagcactt aaatcctcat cctgggcata tatagacttc tgtaattctt
     2821 gtctccctag tactcatcac agagctttcc tcccatggca gggcataacc cacagttgcc
     2881 ggccatttgt tgtttcaggt ggtgaaatac agactaagag cagggacttg cctccagtca
     2941 agaagcttcc agaaaaggag catgggaaga tatgccacct gagggaagac attgcccaga
     3001 ttcctacaca tgcagaagct ggtgaacagg agggcaggtt acaaagaaag cagaaaaatg
     3061 ccatagggag taggcgacat tattgccatg aatgtggaaa gagttttgct caaagttcag
     3121 gcctgactaa acacaggaga atccacactg gtgagaaacc ctatgaatgt gaagactgtg
     3181 gaaagacctt cattgggagc tctgcccttg tcattcatca gagagtccac actggtgaga
     3241 agccatatga gtgtgaagaa tgtggtaagg tcttcagtca cagctcaaac cttatcaaac
     3301 accagagaac ccacactggg gagaagccct atgagtgtga tgactgtggg aagaccttca
     3361 gccagagctg cagcctcctt gaacatcaca aaattcatac tggggagaag ccataccagt
     3421 gcaatatgtg tggcaaagcc tttaggcgga attcacatct cctgagacat cagaggattc
     3481 atggtgataa aaatgttcag aatcctgagc acggggagtc ctgggaaagt cagggtagga
     3541 cggaaagcca gtgggaaaat actgaggctc ccgtgtctta taaatgtaat gagtgtgaga
     3601 gaagtttcac acggaataga agtcttattg aacatcagaa aatccacact ggtgagaaac
     3661 cctatcagtg tgacacatgt ggaaaaggtt tcactcgaac ttcatacctt gttcaacatc
     3721 agagaagcca tgtagggaaa aaaactcttt cacaggggat tccaactact gcaagcgaga
     3781 atctttattt tcagggcgcc gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc
     3841 agcacatgga cagcggaggt ggaggttcga gcggtccctc gggttcgtcg agcctggaag
     3901 ttctgttcca ggggcccctg tcgtcgagcg gtccctcggg ttcgatggtg agcaaaggcg
     3961 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4021 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4081 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4141 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     4201 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4261 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4321 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4381 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     4441 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4501 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4561 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtccttaag gagttcgtga
     4621 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgatct agagtcgagt
     4681 taattaagaa ttccgccccc ccccctctcc ctcccccccc ctaacgttac tggccgaagc
     4741 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     4801 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     4861 ctttcccctc tcgccaaagg aatgcagggt ctgttgaatg tcgtgaagga agcagttcct
     4921 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     4981 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     5041 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     5101 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     5161 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     5221 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     5281 cacaacctgt tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc
     5341 ttacataaac agtaatacaa ggggtgttat gggatcggcc attgaacaag atggattgca
     5401 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5461 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5521 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5581 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5641 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5701 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5761 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5821 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5881 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5941 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6001 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6061 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6121 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagtctgacc
     6181 catggtattc atgccaacat tggtgctcat ttgtcactga actgaagcca ccctggagcc
     6241 cttaatggtt acagaaattg tggaccaggg ctttatttac cactatacat catcgggtat
     6301 tagaaaatgt agtctggctg agacgatgag ttatcttcta cattctagaa agtgattgga
     6361 agaagtcatt tgataagagg ccatgggaga gtggtctgga agttgtagga cctaaaatag
     6421 tttgttctca cccagtgtat ttaaatggcc ttctggactt gttaattgcc ctcagccagc
     6481 actttaggac atctggttgg atggctctga tgtggggggc tgctgctctg accattcatt
     6541 ttgcctctaa tgaatctttc acaaattggt cagatccatg aggtctttta ctcttctttg
     6601 tgccttgtat gataggtgct aataaacatt tattaaatgt accagtaaaa tggcctttgt
     6661 accaaaaatt tgatgttgtc tatatttctg agaaatttct aacttatgcc atttgaattt
     6721 atgtattatt ctctgaggtt ctagattctg ggccaatcta gtgtatttgt tgccacgtac
     6781 atactgtttt cagaattggg gtgggggaat ccatatttct ctaccatgat agtagagatg
     6841 agaagaatgc cttaaacagc ttctttatca ttaagacact gtgagattgc tccatccata
     6901 gaactccact gtctggtcaa tgttaattat gttagtttac attaattttg ctactcttta
     6961 gggggagtct ttcacattca ggtaaataaa aatcaactta ctgagttaga tcaaaatgat
     7021 tattatgaat tttgaacaca ttctgtaaca cttaccatct tgtttcttac cttttagtta
     7081 tcttgggccg accaagtgct aagacgtcat ctgtgtccct cttccc
//
