LOCUS       sequence-211749-        7010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZBTB1 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7010)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7010)
  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: pZBTB1-donor
FEATURES             Location/Qualifiers
     source          1..7010
                     /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             3653..3673
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3686..3730
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3770..3793
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3821..3823,3824..3826,3827..4540)
                     /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(3866..3887)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4127..4146)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4578..5151,5152..5154,5155..5163)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4744..4761)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4971..4990
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5245..6048
                     /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(5308..5327)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5918..5937
                     /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 tcggtacccg aaaacacagg aatgtctagg gccaagggat
     2701 gaccaaaatt taaaagatta tttcatatca gagtcaggga tattaataat tttatgtgag
     2761 aaaatttgca gtttgttttc tcctatttga ggaaggaaca gttaattata cagcttaaaa
     2821 agcccctact tacttaaaaa gttaacatgt ttttattctc cttaaactaa ctgctcatat
     2881 caaacacaat caggagagct catttttaga tgatagaaac attaattcta agcatccaga
     2941 ttatgaacca tctttttttt caaatgctta tgtttaaaaa attttaaaaa aattaaggat
     3001 cttattatga gggaaaagca ctgttgccaa aaatataatt ttactcagaa tcactagggc
     3061 agattaggag gcttataagc ctaggaattt gcaggaaaac ccctcctctt cttttgtgcg
     3121 tgtatgtgtg tgtgcgtgta tgcttcattt tcatttgcag agtggctgaa gacagtaaaa
     3181 agaatatgaa ctttaaaaaa attgttagat ttatgtttca atcagagatt tgtcattaac
     3241 aagttaaagt acattgtaaa atcattttat ttgaaggaaa acacttctta actgcttcct
     3301 tcctccctac tttcctctat ttgtctgacc cctatctttt tttatgttat atatttatat
     3361 gtcctgctga tagattaata agaacatctt atatttgaat ggtacgagtt acattcggga
     3421 aataattaaa tttaaattct aggggctatg cttgtgtttc tattctctgt tctgttaaca
     3481 gtgtgttggc ctagtgccaa agccaagaaa ttctgatcta agaattatgt tgtatcttgt
     3541 agagcagtca acttgtaata ttatttttct tgagttttgt ctttttctgt gtggcagtgg
     3601 tgaaataggg ccttctaaac ctgtggagaa ggggattcca actactgcaa gcgagaatct
     3661 ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca
     3721 catggacagc ggaggtggag gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct
     3781 gttccagggg cccctgtcgt cgagcggtcc ctcgggttcg atggtgagca aaggcgagga
     3841 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3901 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3961 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     4021 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4081 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4141 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4201 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4261 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     4321 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4381 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     4441 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc
     4501 cgccgggatc actctcggca tggacgagct gtacaagtaa tgatctagag tcgagttaat
     4561 taagaattcc gccccccccc ctctccctcc ccccccctaa cgttactggc cgaagccgct
     4621 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4681 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4741 cccctctcgc caaaggaatg cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     4801 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     4861 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     4921 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     4981 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     5041 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     5101 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     5161 acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact gtctgcttac
     5221 ataaacagta atacaagggg tgttatggga tcggccattg aacaagatgg attgcacgca
     5281 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5341 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5401 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5461 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5521 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5581 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5641 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5701 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5761 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5821 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5881 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5941 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6001 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt ctgagagatc
     6061 tgaatttgga gaggacatgg ggaagaatca atcttcaaca gataatcttt aagtacagac
     6121 ccaggaatat cagatctgaa gtggcaccat cttttctgtc ttcctgacaa acttccatca
     6181 acccaaaaag ttccagttgc aaagatccaa gaaaactcag taaagatcac tttggaatca
     6241 ctttgttgta acaaaattga ttcatggtga ctctaatatt tgtgtaaatc tgctgaagaa
     6301 aagtaactag aaaattcata caacatcagt ggttacagct atactaaaat aaaactttat
     6361 atgctaatta tcttcaaggg tctttctaaa ataaaagggg ttgttaggct actgcaaata
     6421 taaagagaca taatcacaaa tctatttata tagatataaa taattctaaa cttaaaatca
     6481 ctagaattat agcttaaata ttatgaaatg tacttagtca acttaaaaac ttacaccaaa
     6541 aatctgtagt ttagagtgta agaagtaatg ggtacagaga aaaaaaattt aagaccaata
     6601 ttttaacagg ataatttggt tattactcac tatagtaaaa cacttgccct tcttcctttt
     6661 ccactgtata tgccagtcac aatcttgatc atttatatga atgaaactcc cttttaaacc
     6721 tgttaacttg aaggagaaca gaaatggtat tacttcaata gtggtagatt tccacaaatc
     6781 agacacatta ttttcttctg aacttggcaa tataacctaa taaggcagaa taagtttctg
     6841 ttaaaatact tattctcaat atttatgtat aaaccatgct tctcagcttt caaatttcaa
     6901 gataaaattt aagtattcag taaaatgatg gttaacatgt tttaaaaaat ctttaatgga
     6961 gaagttaaat atttggtgag attttagtct ggctataaat gctttgtttg
//
