LOCUS       sequence-230652-        6676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZBTB12 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6676)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6676)
  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: pZBTB12-donor
FEATURES             Location/Qualifiers
     source          1..6676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..36)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(19..35)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(28..50)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(244..263)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     363..385
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(423..441)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        509..613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(614..682,683..1474)
                     /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     complement(832..851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1645..2233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2134..2153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2387..2404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2521..2542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2557..2562,2563..2580,2581..2587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2595..2611
                     /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)."
     primer_bind     2600..2622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2619..2635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2619..2635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2678..2689)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3533..3553
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3566..3610
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3650..3673
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3701..3703,3704..3706,3707..4420)
                     /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(3746..3767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4007..4026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4458..5031,5032..5034,5035..5043)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4624..4641)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4851..4870
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5125..5928
                     /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(5188..5207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5798..5817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
       61 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      121 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
      181 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
      241 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
      301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
      421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2401 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2461 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2521 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2581 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2641 ttacgccaag cttcctccac tcctgcggcc agtgaagctg gagttcccac tggatgaaga
     2701 cttggagctg aaagccgagg aagaggatga ggatgaggat gaggacgtgt ctgacatctg
     2761 catcgtcaag gtggagtcgg ccctggaggt ggcacaccgg ctcaaacccc ctggaggcct
     2821 gggagggggt ctgggcattg gaggctccgt gggtggccac cttggggagc tggcccagag
     2881 cagcgttccc cccagcactg tagccccacc gcagggtgtg gtgaaggcct gctatagcct
     2941 gtcggaagat gcagaagggg agggcctgct gttgattccc ggaggccggg ccagcgtggg
     3001 ggccacctcg ggcctggtgg aagcagcagc ggtggccatg gctgcccggg gggcgggggg
     3061 cagcctgggg gcggggggca gccggggacc cctgcctggg ggcttctcag gtggaaaccc
     3121 cttaaagaac atcaagtgca cgaagtgccc ggaagtgttc cagggcgtgg agaagctggt
     3181 cttccacatg cgggcgcagc acttcatctt catgtgccct cgctgtggca agcagttcaa
     3241 ccacagcagc aacctcaacc gccacatgaa cgtgcatcgt ggtgtcaagt cacactcgtg
     3301 cggcatctgc ggcaagtgct tcacacagaa gtccaccctt cacgaccacc tcaacctgca
     3361 ctcgggagcg cggccctacc gctgctccta ctgcgacgtg cgcttcgccc acaagcctgc
     3421 cattaggcgg cacctcaagg agcaacacgg caagaccacg gccgagaacg tgctggaggc
     3481 cagtgtggcc gagattaacg tcctcatccg cgggattcca actactgcaa gcgagaatct
     3541 ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca
     3601 catggacagc ggaggtggag gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct
     3661 gttccagggg cccctgtcgt cgagcggtcc ctcgggttcg atggtgagca aaggcgagga
     3721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3781 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     4201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     4321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc
     4381 cgccgggatc actctcggca tggacgagct gtacaagtaa tgatctagag tcgagttaat
     4441 taagaattcc gccccccccc ctctccctcc ccccccctaa cgttactggc cgaagccgct
     4501 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4561 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4621 cccctctcgc caaaggaatg cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     4681 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     4741 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     4801 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     4861 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     4921 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     4981 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     5041 acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact gtctgcttac
     5101 ataaacagta atacaagggg tgttatggga tcggccattg aacaagatgg attgcacgca
     5161 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5221 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5281 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5341 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5401 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5461 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5521 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5581 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5641 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5701 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5761 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5821 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5881 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgacc gcgcaggcgt
     5941 ggaggccagg aggctggggc ccctgggctg cgtggaaaaa gggctctttg gcccaggaga
     6001 attggggggt ggggggtctg gggcagaaag gtaagagtgg gaggctgagc agatgcacac
     6061 atcctgagag agggaagatg attccttgga gagacttgct cttgagagtg caagaatctg
     6121 gagctgggaa aagggttctt ggaggccagg ggaatacggg gtcccagaga aagatttcct
     6181 tctcttagaa gtgcatgtat atgtggaggg agggaaaagg gtcctataga atgagggaag
     6241 acaaaaatgt tttattcctg gctaaggctg cccaggggaa ggttctgaca tttctcgaga
     6301 taagaagggt tggggggtgg gtgggaacag ggagggaatt tggcaggagg acatgctcct
     6361 gcttagaata gacctggagt gagggagtgg taaggggaaa agattactga gtcccagaaa
     6421 aggaagtggg agacagttct tgcatgctgg ggaaatgaga ctgcgagtat gaggcttgct
     6481 gaaatttgag ggagaacaaa gtaagacgcc aggtaagaga acgtggggtg cgcatcctgg
     6541 gtagatattg ggaagtgggg gtacaatagc aagaaacaat ggatagtcat gggggtgggg
     6601 gggatagggc acttagaaaa gaaagaacac aagaagggct gttgggaggg agggagagga
     6661 gtagagacat acagta
//
