LOCUS       sequence-228569-        7131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZBTB40 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7131)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7131)
  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: pZBTB40-donor
FEATURES             Location/Qualifiers
     source          1..7131
                     /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             3714..3734
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3747..3791
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3831..3854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3882..3884,3885..3887,3888..4601)
                     /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(3927..3948)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4188..4207)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4639..5212,5213..5215,5216..5224)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4805..4822)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5032..5051
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5306..6109
                     /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(5369..5388)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5979..5998
                     /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 acggagaagt agctgggctt ggcttcctca
     2701 cttgttcctg ctctctgctc ttcactgatt ctcaagagca aggtggctgt ggcctctagt
     2761 ttaaaaatat caagggcatc tagctcttct gggatctatg ggttagaaac atcaactggg
     2821 aggcttaggc gggaattgga tatgcctaac cacagactca ttctctctca gttggaaagg
     2881 tggtctctaa cagattatgc agatttggaa gggacatagg tgggctgtgg aggagagaag
     2941 ggggcactgg ccccgccagg ctggtgagct gaatgaatgc gtggagtcac agttcatcca
     3001 gtcttcctcc taagctcggt cactgtgtgg cctcctggga aggctgtgag cctgcgagag
     3061 gtccagagag gctgccccgg gcccacctgg gcctgtggtg gatcaatccc attcccatgt
     3121 tccaggcatg agaccagctg gcttcagtgg cattcctctc acatggcact cacctctgca
     3181 gtgtcatccc aagacaggaa agcccagtgt gaaaaacaga gccagtgttc gctggtgtat
     3241 aatcatagag tccgggcgcc ccgagatggc gtactgagca aaattatagc aatttagtgc
     3301 acagttattt aaattgtttg ctggccttct cttttgcatc gccagtgggt tagacttgcc
     3361 tcttcccatt aggcacatgt aagtgctctc ctcaggtgaa aacataaata tcatcattac
     3421 agcatgagat gaaaaccatg taaatcaggg agccaactgt gaacactgcc cagagcagcc
     3481 tcacggtctt tctctttcag gtgatccaaa ccccagagcc ggtggccccg acagagcagg
     3541 tgatcacttt ggaggagacc cagcttgccg ggtcgcaggt gtttgtgacg ttgccagatt
     3601 ctcaggcatc tcaggccagc tctgagctcg tggcggtgac tgtggaggac ttgctggatg
     3661 gcacagtgac gctgatctgt ggtgaggcca aagggattcc aactactgca agcgagaatc
     3721 tttattttca gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc
     3781 acatggacag cggaggtgga ggttcgagcg gtccctcggg ttcgtcgagc ctggaagttc
     3841 tgttccaggg gcccctgtcg tcgagcggtc cctcgggttc gatggtgagc aaaggcgagg
     3901 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3961 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     4021 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     4081 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4141 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4201 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4261 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4321 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4381 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4441 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4501 ccctgagcaa agaccccaac gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg
     4561 ccgccgggat cactctcggc atggacgagc tgtacaagta atgatctaga gtcgagttaa
     4621 ttaagaattc cgcccccccc cctctccctc ccccccccta acgttactgg ccgaagccgc
     4681 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     4741 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     4801 tcccctctcg ccaaaggaat gcagggtctg ttgaatgtcg tgaaggaagc agttcctctg
     4861 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     4921 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     4981 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     5041 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     5101 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     5161 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     5221 aacctgttac attgcacaag ataaaaatat atcatcacga acagtaaaac tgtctgctta
     5281 cataaacagt aatacaaggg gtgttatggg atcggccatt gaacaagatg gattgcacgc
     5341 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5401 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5461 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5521 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5581 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5641 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5701 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5761 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5821 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5881 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5941 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6001 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6061 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag tctgagcagc
     6121 ctttcatccg gcagagcctt cctgcgtttg cagcagagag gaggccccac agcttgccct
     6181 ttgccctcca tccctggctg tcctgagtgg tgagcatctt agcttagcac caaccaacac
     6241 agtctcacct agaaaacaga tggaagcttc gttgttctca tagaaccaac agcatctgag
     6301 ccctcaacac caacagcacc atcctctgta gcagacaggc ctccctcccc acaggcccgc
     6361 tgctgcggcc tctatgacac tgtccatccc caagtgacat gtggcttcag aagtagagtc
     6421 tgaaagagca gtgggatggg agctggtgac cagggtaccc caggaggcat gatgtgccga
     6481 cagcgctcag tgggcagaat ggcagttaga tatgggactt ggcccacagt gggggctgca
     6541 aatgctgcca ctgcctctgg ccatttaaag tgagaggggc accaacagac aattcgggga
     6601 ccttaggccc cttcctgagg cacacttggc cccttcctcg gtcctatcat cctaccctct
     6661 gctggggttc cccctccacc cagtggccac gcccagcacc ctattgatgg ctataggaca
     6721 ggtagccctc atttccatgc ctgataaccc cttgtcagtt gtcagctctt cccaaaacaa
     6781 agctctggag tttctgctgg aagtgtttaa tgtgaggcat cagctgctag acagtgtctg
     6841 cctttccagg acattcttct tggtattcct tacctgaagc ctggttccca cagctcttcc
     6901 gtgtcatctt catccccttc ctttaatcaa aggctgaaat ctctgcctga cctggcagcc
     6961 tggctccctc tgggtcatat ggtgcagact gtgaccagca ccaggcaggc agtcctgtct
     7021 gcgtgtggag gagcagccgt gactgcccgt ggctctgctg ccgcccactc cctgccttgt
     7081 gagtggcctg ctgcctcacc tcccgcaggc cgccacactt attgcaggtc a
//
