LOCUS       sequence-211717-        7267 bp    DNA     circular SYN 15-APR-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF347 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7267)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pZNF347-donor
FEATURES             Location/Qualifiers
     source          1..7267
                     /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             3881..3901
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3914..3958
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3998..4021
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4049..4051,4052..4054,4055..4768)
                     /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(4094..4115)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4355..4374)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4806..5379,5380..5382,5383..5391)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4972..4989)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5199..5218
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5473..6276
                     /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(5536..5555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6146..6165
                     /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 cctagctatt catctggtaa ttcacaccgg
     2701 agaaaagcct tacaaatgtc atgaatgcgg caaggtcttt aggcgtaatt cacaccttgc
     2761 aaggcatcag ctaattcata ctggagagaa accttataag tgtaatgagt gtggcaaagc
     2821 ctttagagca cattcaaacc taactaccca tcaggtcatc catactggag aaaagcctta
     2881 caaatgtaat gaatgtggca aggtcttcac tcaaaattca caccttgcaa atcatcaaag
     2941 aattcatact ggagtgaagc cttatatgtg taatgagtgc ggcaaagcct tcagtgtgta
     3001 ttcaagccta actacccatc aggtcatcca tactggagaa aaaccttaca aatgtaatga
     3061 gtgtggcaag gtcttcactc agaattcaca ccttgcaaga catcggggaa ttcatactgg
     3121 agagaaacct tataaatgta atgaatgtgg caaggtcttt aggcataatt catacctttc
     3181 aaggcatcag cgaattcata ctggagagaa accttacaag tataatgagt atggcaaagc
     3241 ctttagtgaa cattcaaacc taactaccca tcaggtcatc catactggtg aaaaacctta
     3301 caaatgtaat gagtgtggca aggtcttcac tcagaattca caccttgcaa gacatcggag
     3361 agttcatact ggaggtaaac cttaccagtg taatgaatgt ggcaaagcct ttagtcaaac
     3421 atcaaagctt gcaaggcatc agagagttca tactggagag aaaccatatg agtgtaatca
     3481 gtgtgggaaa gcctttagtg tccgttcaag cctaactacc catcaggcaa tccatactgg
     3541 aaaaaaacct tacaaatgca atgagtgtgg gaaggtcttc actcagaatt cacaccttgc
     3601 aagacatcgg ggaattcata ctggagagaa accttacaag tgtaatgagt gtggcaaagc
     3661 ctttagtcaa acttcaaaac ttgcaaggca tcagagaatt cataccggag agaaaccata
     3721 tgagtgtggg aaacccttta gtatctgttc aagcctaact acccatcaga caatccatac
     3781 tggtgggaaa ccttacaaat gtaacgtgtg gaaagttcta aagtcagagt tcaaaccttg
     3841 caagccatca cagaattcag ggattccaac tactgcaagc gagaatcttt attttcaggg
     3901 cgccgccaaa ttcaaagaaa ccgctgctgc taaattcgaa cgccagcaca tggacagcgg
     3961 aggtggaggt tcgagcggtc cctcgggttc gtcgagcctg gaagttctgt tccaggggcc
     4021 cctgtcgtcg agcggtccct cgggttcgat ggtgagcaaa ggcgaggagc tgttcaccgg
     4081 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     4141 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     4201 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     4261 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4321 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     4381 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     4441 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     4501 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     4561 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4621 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     4681 ccccaacgag aagcgcgatc acatggtcct taaggagttc gtgaccgccg ccgggatcac
     4741 tctcggcatg gacgagctgt acaagtaatg atctagagtc gagttaatta agaattccgc
     4801 ccccccccct ctccctcccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     4861 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     4921 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     4981 aaggaatgca gggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     5041 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     5101 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     5161 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     5221 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     5281 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     5341 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac ctgttacatt
     5401 gcacaagata aaaatatatc atcacgaaca gtaaaactgt ctgcttacat aaacagtaat
     5461 acaaggggtg ttatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc
     5521 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5581 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5641 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5701 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5761 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5821 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5881 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5941 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6001 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6061 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6121 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6181 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6241 atcgccttct atcgccttct tgacgagttc ttctgagtct ggagagaaac cttacaaatg
     6301 aaggtggtaa cttttgcagt tagaattcat tcctttgaca acgtcagaga atttatgcct
     6361 gagagattcc ttgaaaatgc aatgactggc tgggtgcagt ggctcatgcc tgtaatccca
     6421 acactttggg agtccgaggc aggtggatca cctgaggtca ggagttcaag accagcctga
     6481 ccaacatggc gaaaccccat ttctactaaa agtacacaat tagctaggcc tggtggcgtg
     6541 cgcctgtaat cccagctata taggaggctg aggcaggaga atcacttgaa cccaggaggc
     6601 ggaggttgca gtgagccaag atcgtgccaa tgcactccat cctgggcaaa aagagcaaaa
     6661 ctccatctta aaggaaaaaa aaaaaatgca atgactgtgg caaactcctc atgagttcaa
     6721 gtattaatag acatccaaga gtccataata atgagaaatc acataaatgt actgtgtgtg
     6781 acagaggctt tattcaggcc tcacagcttt gcaggcatta aaatctacgt atttgatgga
     6841 accacacaaa tgtaatgtgt aagagcatcc tgaagctctt actctaagac tgtaactgta
     6901 ggtgaggatt catacaaaga gtgactcagt ctctagttct ttgattcact tttaaaatga
     6961 taaactgcac aataattaca cgttccaata tggtaaaact taagacatta tatatttcaa
     7021 aggtcaaagg acatttggaa atggctactt accttcaggt tataaaatct tttattccat
     7081 tgtttgaggt ttctcaaaaa ggctactgtt tgtgactttc aacctgaact atggatctgt
     7141 ggctattttg catccttaag cacctttcgt ggtgcctatc tgggaaagaa ccacagtatg
     7201 gaaaggctta cttcattaca tgaggatcat ttttatccat gtttattgaa gaaggaggac
     7261 tctgcct
//
