LOCUS       Exported                7249 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF84 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF84-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7249)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7249
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(82..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(1195..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             3797..3817
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3830..3874
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3914..3937
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3965..4684
                     /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(4010..4031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4271..4290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4722..5307
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4888..4905)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5115..5134
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5389..6192
                     /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(5452..5471)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6062..6081
                     /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 aggaacaaaa ccctttggat gtagtgattg
     2701 tagaaaagcc ttctttgaga agtcagagct tattagacat cagacaattc atactggaga
     2761 gaaaccctat gaatgcagcg agtgtaggaa agcatttaga gagaggtcga gtctcattaa
     2821 tcatcagaga acacatacag gagagaaacc tcatggatgc attcagtgtg ggaaggcctt
     2881 ctcccagaag tcacatctca tatcacatca gatgacacac acaggagaaa aaccctttat
     2941 atgcagtaaa tgtgggaaag ccttcagcag gaaatcacag ctcgttagac atcagagaac
     3001 tcatacggga gaaaaaccgt atgaatgcag tgagtgtggg aaagctttca gtgaaaaatt
     3061 aagtctcact aatcatcaaa gaattcatac aggagaaaaa ccatatgtat gcagtgaatg
     3121 tgggaaagcc ttttgtcaga agtcacatct catatcacat cagaggacac atacagggga
     3181 gaaaccctat gaatgcagtg aatgtgggaa ggcctttggt gagaagtcaa gtcttgcaac
     3241 tcatcagaga actcatactg gagaaaaacc gtatgaatgc agggactgtg aaaaagcttt
     3301 ctcccagaaa tcacagctaa atacccatca gagaattcac actggagaga aaccctatga
     3361 atgcagtctt tgtaggaaag ctttttttga gaagtcggag ctaattagac atctgagaac
     3421 tcatacagga gaaaaacctt atgaatgcaa tgaatgtaga aaagccttca gggagaagtc
     3481 aagtctcatc aatcatcaga gaatacatac aggagagaag ccttttgaat gcagtgagtg
     3541 tggcaaagct ttctctcgga agtcacacct tataccacat caaaggacac atacgggtga
     3601 gaaaccctat ggatgcagtg aatgtaggaa ggccttctct cagaagtcac agctggttaa
     3661 tcatcagaga attcatacag gagagaagcc ttatcgatgc attgaatgtg ggaaagcttt
     3721 ctcacagaag tcacagctca tcaatcatca gagaactcat acagtaaaaa aatccgggat
     3781 tccaactact gcaagcgaga atctttattt tcagggcgcc gccaaattca aagaaaccgc
     3841 tgctgctaaa ttcgaacgcc agcacatgga cagcggaggt ggaggttcga gcggtccctc
     3901 gggttcgtcg agcctggaag ttctgttcca ggggcccctg tcgtcgagcg gtccctcggg
     3961 ttcgatggtg agcaaaggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4021 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4141 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     4201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     4321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     4381 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     4441 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     4501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     4561 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     4621 ggtccttaag gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4681 gtaatgatct agagtcgagt taattaagaa ttccgccccc ccccctctcc ctcccccccc
     4741 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     4801 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     4861 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcagggt ctgttgaatg
     4921 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     4981 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     5041 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     5101 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     5161 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     5221 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     5281 aaacacgatg ataatatggc cacaacctgt tacattgcac aagataaaaa tatatcatca
     5341 cgaacagtaa aactgtctgc ttacataaac agtaatacaa ggggtgttat gggatcggcc
     5401 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5461 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5521 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5581 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5641 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5701 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5761 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5821 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5881 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5941 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6001 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6061 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6121 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6181 gagttcttct gagtctagga atacagttaa tagtagtctt tgacagatca tcttggactt
     6241 caggaaatgc aattatgata acgtttgtag acagtcacgt catgttaggt gtctgtactc
     6301 catgaggatg agaactctaa atgaggtggt gtatggaaag ccgatcataa ttcatagagt
     6361 agagtgaacc tatgactgca gtggatctca aaaactttta aaaccataga caagccttat
     6421 agagtagaac attcacagca aagaagaatc ctgtgaatgt ccaaaagcct tccagaagtc
     6481 aagtctctta agctattaga aatattccca ctggggatga gggaaaaccc catgaatgcg
     6541 ggaaatgagg caatattttt aagaaatgac agttcattgt acataagaaa atgctcttag
     6601 gaatgaagtt ctatgaaagt actaaatatg ggacagtgca acaagtaacc agactatttt
     6661 gtattttgga gaattcatat tatggagaac ctaacaattt aaagacactg ggaacacttg
     6721 cccctcagta tagtactgtc aagggaagcc atacactttt tgtagacatg ggtaccaaaa
     6781 atacccaatt ctaagtggtt gacagatgtt cactttgaag tgtgaagttt taaaaatacg
     6841 tgaataaatt ggttattgaa acatctaaga catttcttag tggtatttgt ggcttatctt
     6901 ctagtgtcac gtatgttttg tgttttggac ttacctataa tttaatgtaa ttgggagtat
     6961 gaaatgcata tgccttttag tgcctctgat gtctgtgaag acaacactac acaccactgg
     7021 tttttatttt ttaacaataa tataaaaaat ggaaactcat gtaacatgac tacatgaata
     7081 agtaccttaa gtgataaccc gtttctttta acttatagac atacataagg ggtcttttgt
     7141 ttttatggac ctttctattc agtaactgaa gccaacattg tgtaagtctt ctgcatcctc
     7201 agagtctgag taacagttta tataaaataa atatgcaaag gcaggaacc
//
