LOCUS       sequence-229653-        7074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF589 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7074)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7074)
  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: pZNF589-donor
FEATURES             Location/Qualifiers
     source          1..7074
                     /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             3702..3722
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3735..3779
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3819..3842
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3870..3872,3873..3875,3876..4589)
                     /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(3915..3936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4176..4195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4627..5200,5201..5203,5204..5212)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4793..4810)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5020..5039
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5294..6097
                     /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(5357..5376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5967..5986
                     /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 aaaatatagg tcctgcacag cacatcttgg
     2701 ggagaaagac tttaatcatt gagaatgatt aatgggccag gtcttatcat aaaacaagag
     2761 cccagtcagc ccttcctatg actcttctga ctggaaactt tctttcttca gcagaatcaa
     2821 agccagaagt ccatacctgc ccttcttgcc ctctggcctt tggcagtcag cagttcctca
     2881 gccaagatga gctacacaat catcctattc caggtttcca tgcaggaaat caactccacc
     2941 caggaaatcc ctgcccagag gatcagccac agtcacaaca tccttctgat aaaaatcaca
     3001 ggggggctga agcagaagat caacgagtgg aaggaggcgt cagacccttg ttttggagta
     3061 caaatgaaag gggggcttta gtgggtttct ctagcctgtt ccagagacca ccaataagct
     3121 cttggggagg caacagaata ttagagatac agctcagtcc agcccagaat gcaagctctg
     3181 aggaagtaga cagaatttcc aagagggcag aaaccccagg gtttggagca gtcacgtttg
     3241 gggagtgtgc actagctttt aaccagaagt caaacctgtt cagacagaag gcagtcacag
     3301 cagaaaaatc ttcagacaaa aggcagtcac aggtgtgcag ggagtgtggg cgaggcttta
     3361 gcaggaagtc acagctcatc atacaccaga ggacacacac aggagaaaag ccttatgtct
     3421 gcggagagtg tgggcgaggc tttatagttg agtcagtcct ccgcaaccac ctgagtacac
     3481 actccgggga gaaaccttat gtgtgcagcc attgtgggcg aggctttagc tgcaagccat
     3541 acctcatcag acatcagagg acacacacaa gggagaaatc gtttatgtgc acagtgtgtg
     3601 ggcgaggctt tcgtgaaaag tcagagctca ttaagcacca gagaattcac acgggggata
     3661 agccttatgt gtgcagagat gggattccaa ctactgcaag cgagaatctt tattttcagg
     3721 gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg
     3781 gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc
     3841 ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg
     3901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3961 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4081 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4141 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4261 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4321 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4381 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4441 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4501 accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca
     4561 ctctcggcat ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg
     4621 cccccccccc tctccctccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     4681 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     4741 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     4801 aaaggaatgc agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     4861 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     4921 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     4981 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5041 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5101 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5161 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa cctgttacat
     5221 tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa
     5281 tacaaggggt gttatgggat cggccattga acaagatgga ttgcacgcag gttctccggc
     5341 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5401 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5461 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5521 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5581 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5641 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5701 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5761 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5821 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5881 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5941 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6001 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6061 catcgccttc tatcgccttc ttgacgagtt cttctgagtc tgaggccgag gctttgtaag
     6121 gagatcatgt ctcaacacac accagaggat acattcagat gagaagcctt ttgtttgcag
     6181 agagtgtggg cgaggctttc gtgctaaatc aactctcctc ctacaccagt ggacacattc
     6241 agaggtgaaa cctcacgtgt gtgaggagtg tgggcatgga tttagccaga agtcgtcgct
     6301 caaatcacat cggagaacac actcagggga gaagccttat gtgtgtgggg aatgtgggcg
     6361 gggatttagc cggaggatag tcctcaatgg acactggagg acacacacgg gagagaagcc
     6421 ttacacgtgc tttgagtgtg ggcgaaactt tagcctcaag tccgctctta gtgtacatca
     6481 gaggatacac tctggggaga agccttatgc atgcacggag tgtgggcaag gctttatcac
     6541 gaaatcacag ctcatcagac accagaggac acacacagga gaaaagcctt atgtctgcgg
     6601 agagtgtggg cgaggcttta tagctcagtc aaccctccac taccaccgga gtacacactc
     6661 caaggaaaaa ccttatgtgt gcagccagtg tgggcgaggc ttttgtgata aatcaactct
     6721 cctcgcacac gagcagacac attcagggga gaagccttat gtgtgtgggg aatgtgggcg
     6781 gggatttggc cggaagatac tcctcaacag acactggagg acacacacag gagagaaacc
     6841 ttacgcatgc atcgagtgtg ggcgaaactt tagccacaag tccactctca gcttacatca
     6901 gaggatacac tcgggggaga agccttatgc atgcgtggag tgtgggcaaa gctttaggag
     6961 aaagtcacag ctcatcatac accagaagat acactcgggg aaaagcttta gaggtgcaag
     7021 gagtgaggat gtgattttag caacaagtca gccatcagcc acaccagcgg aaat
//
