LOCUS       sequence-211763-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF416 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7377)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7377)
  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: pZNF416-donor
FEATURES             Location/Qualifiers
     source          1..7377
                     /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             3888..3908
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3921..3965
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4005..4028
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4056..4058,4059..4061,4062..4775)
                     /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(4101..4122)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4362..4381)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4813..5386,5387..5389,5390..5398)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4979..4996)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5206..5225
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5480..6283
                     /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(5543..5562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6153..6172
                     /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 agaagccaaa caaaatcagc aaatgtgagg
     2701 aggcctttca tgttggaata agtcattaca agtggagtca atgcaggaga gagtccagcc
     2761 acaaacacac tttttttcac cctagagtct gcactggaaa aaggctttat gaatctagca
     2821 aatgtgggaa agcctgctgc tgtgagtgct cccttgttca gctgcaaaga gtccaccctg
     2881 gagaaaggcc ttatgagtgc agtgaatgtg ggaaatcttt tagccaaacc tctcatctga
     2941 atgatcatcg gagaatccac actggagaaa ggccttatgt gtgtggtcag tgtgggaaat
     3001 catttagcca aagagccacc ctcattaaac atcacagagt tcacactgga gaaaggcctt
     3061 acgagtgtgg tgaatgtggg aaatctttta gccaaagttc caaccttatt gaacattgca
     3121 gaattcacac tggagaaagg ccttatgagt gtgatgaatg tggaaaagcc tttgggtcca
     3181 aatccactct tgttcgacac cagagaactc acacaggaga aaagccatat gagtgtggtg
     3241 aatgtgggaa attattcaga caaagcttca gccttgttgt acaccagaga attcacacta
     3301 cagcaaggcc ttatgagtgt ggccagtgtg ggaaatcatt tagcctaaag tgtggcctca
     3361 ttcagcacca gttaattcac agtggagcta ggccctttga gtgtgatgag tgcggaaaat
     3421 cctttagcca aagaaccacc ctcaataaac accacaaagt tcacactgca gaaaggcctt
     3481 atgtatgtgg ggaatgtggg aaagctttta tgttcaaatc taaacttgtt aggcaccaga
     3541 gaactcacac tggagaaagg ccttttgagt gcagtgaatg tgggaaattt tttagacaaa
     3601 gctataccct cgttgaacac cagaaaattc acactggatt aaggccttac gactgtggac
     3661 agtgcgggaa atcctttatc caaaagtcta gcctcattca acaccaagtg gttcacacag
     3721 gagaaaggcc atatgagtgt ggcaaatgtg ggaagtcctt tacacaacac tctggcctca
     3781 ttctccaccg aaaatctcac actgtggaga ggcctcgtga cagcagcaaa tgtggaaaac
     3841 cctacagccc aagatctaac attgttggga ttccaactac tgcaagcgag aatctttatt
     3901 ttcagggcgc cgccaaattc aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg
     3961 acagcggagg tggaggttcg agcggtccct cgggttcgtc gagcctggaa gttctgttcc
     4021 aggggcccct gtcgtcgagc ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt
     4081 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     4141 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     4201 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     4261 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     4321 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     4381 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     4441 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     4501 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     4561 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     4621 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     4681 gcaaagaccc caacgagaag cgcgatcaca tggtccttaa ggagttcgtg accgccgccg
     4741 ggatcactct cggcatggac gagctgtaca agtaatgatc tagagtcgag ttaattaaga
     4801 attccgcccc cccccctctc cctccccccc cctaacgtta ctggccgaag ccgcttggaa
     4861 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4921 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4981 ctcgccaaag gaatgcaggg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     5041 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     5101 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     5161 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     5221 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     5281 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     5341 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacctg
     5401 ttacattgca caagataaaa atatatcatc acgaacagta aaactgtctg cttacataaa
     5461 cagtaataca aggggtgtta tgggatcggc cattgaacaa gatggattgc acgcaggttc
     5521 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5581 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5641 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5701 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5761 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5821 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5881 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5941 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6001 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6061 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6121 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6181 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6241 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagtcctct tgaaactcca
     6301 aacctgagaa aagccttaga cctgcaggga atgtgccatg tctttcttca gtgttatggc
     6361 tatagagagc ctttgtgaag gagccgtctg cctaaagtta aacctcatac ttctagacct
     6421 cccgatgggt cagactccca ttagttccag gcatatgtgt cttgcaggag ctcctttgca
     6481 tgttctaaac tgctggaggc ctttgccaga gttaaatcat tagtcaccct tgcatactaa
     6541 ggcgaggcag atacacctgt gttctcccaa cccttggagg aaattccttg agcagtctga
     6601 gcctttaagg ggaattcctt cccttttctg actgattaca gcatacatct gaccagtttt
     6661 ggccagaagg gcccagcctg ggttctgctg gaagctgatg tggaaggctt cccttcatgg
     6721 aaattcttgt tttcacacaa cacttggtag tttttccagc ctccaagtca ccacctagta
     6781 actggctgcc tcacattgct ctggtttgtg tggtaataaa ggccttattt ttaaatctat
     6841 gtgtggtctt agggtttggt ttactgtgtg aggtggctgg aaaacacatt tgggctccta
     6901 cataaaggca tggacagctt ttgtggggcc ccaatgttgc tggcctgtgc aggaaaagta
     6961 aaatgacagt agttcttgtt ctggttttag tccactttgc cttgctgctc aaggcctcct
     7021 aggaaacaat gcaagaattt tgttgtccta atttatggcc tggatcccta cactttctct
     7081 ggaccagagg tcatcctgag gacaagtcgg ctgttatgac aggctcctgt gctttaggga
     7141 ataagatgga tttattgcat tgttctacag gctgtaatgc ccagcactgt tgagacgaag
     7201 ctttttccca gcttctgcaa aggaacctat gtgccctgat gcccatgatt ccatagatgg
     7261 tggtactcaa gctgatggga gctaagcatg ggacataatt ggtacagaaa ttttgggtat
     7321 gattaataga aagtggagga ggtaatagca aaacaaatgt gcctgtggta aaagtgc
//
