LOCUS       Exported                7380 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF257 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF257-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7380)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7380
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1200..1220
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1233..1277
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1317..1340
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1368..2087
                     /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(1413..1434)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1674..1693)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2125..2710
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2291..2308)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2518..2537
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2792..3595
                     /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(2855..2874)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3465..3484
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4781..4797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4781..4797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4794..4816)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4805..4821
                     /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(4829..4859)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4874..4895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5012..5029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5183..5771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5263..5282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5942..6802)
                     /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     6565..6584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6803..6907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6975..6993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7031..7053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7153..7172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7366..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7380..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacccgcat gctttcacaa ctaactcgac
       61 ataagagaat tcatattaga gagaattccc acaaatgtga agaatgtggc aaagccttta
      121 accagtcctc agctcttact cgacataaga tgactcatac tggagagaaa ccctacaaat
      181 gtgaagagtg tggaaaagct tttaaccggt cttcacacct tactcaacat aaggtaattc
      241 atactagaga gaaaccctac aaatgtgaag agtgtggcaa agcctttaac cggtcttcac
      301 acattactca acataagaga attcataata gagagaagcc cttcaaatat gatgaatgtt
      361 gcaaagcctt taagtggtcc tcagctctta ctacccttac tcaacataag agaattcata
      421 ctggagagaa accctacaaa tgtgaagagt gtggcaaagc ctttaaccag tcctcagccc
      481 ttactcgaca taagatgatt catactggag agaaaccctt ccaatgtgaa gagtgtggca
      541 aagcttttaa ccggtcttca caccttactc aacataagat aattcatact aaagagaaac
      601 cctacaaatg tgaagagtgt ggaaaagcct ttaaccggtc ttcacacctt actaaacata
      661 agagaattca tactagagag aaggcctaca aatgtgatga atattgcaaa gcttttaact
      721 ggtcctcagc tcttactacc cttactcagc ataagataat tcatactgga gagaaaccct
      781 acaaatgtga agagtgtggc aaagccttta accggtcttc ataccttatt cgacataaga
      841 taattcatac tggagagaaa ccctacaaat gtgaagagtg tggcaaagcc tttaaccagt
      901 cttcacacct tactcaacat aaaataattc atactgggga gaagccctac aaatgtgaag
      961 aatgtggcaa agcctttaac cggtcttcac acctttctca acataagata attcatactg
     1021 gagagaaacc ctacaaatgt gaagaatgtg gcaagccttt taatcgtttc tcatacctta
     1081 ccgtacataa gagaattcat gctggagaga accccaacaa atatgaagaa tgtggcaaag
     1141 cttgtaacca ttcctcaaac cttactaaac ataattcagg gattccaact actgcaagcg
     1201 agaatcttta ttttcagggc gccgccaaat tcaaagaaac cgctgctgct aaattcgaac
     1261 gccagcacat ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg
     1321 aagttctgtt ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag
     1381 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1441 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1501 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1561 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     1621 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1681 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1741 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1801 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1861 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1921 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1981 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg
     2041 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga tctagagtcg
     2101 agttaattaa gaattccgcc cccccccctc tccctccccc cccctaacgt tactggccga
     2161 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     2221 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     2281 ggtctttccc ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt
     2341 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2401 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     2461 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     2521 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     2581 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     2641 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     2701 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
     2761 tgcttacata aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt
     2821 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2881 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2941 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3001 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3061 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3121 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3181 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3241 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3301 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3361 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3421 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3481 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3541 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtcta
     3601 atggagaaaa accctacaaa tgtgaagaat gtgtcaaagc ttttaactgt tcctcaatcc
     3661 ttactaaaca taagggaatt cataatagag aaaccctaca aatgtgaaga acgtggcctg
     3721 gcttttaaca aatcctcaac atttactaag cataaaataa ttcatgctgg agagaaactc
     3781 ttgaaatgtg atgaatgtgg catagcctct tcccagttct caactcttac taaacatgag
     3841 aacacatgtg gaagataaag cctacaaata tgaagaatgt gacaaggcct ttaaaagttc
     3901 tcaaccctta ttacacataa tttatactgg acagaaatcc taaagtgtga agaatgtcac
     3961 aaagccctta ataagtcctc aattcttaac agatataaga tgtttcatac tggagcgaaa
     4021 ctacaatcct gaatgatgtc acagtgcttt taacaatacc tcaaactttt ctaaacataa
     4081 aaaaaaatta tactagtgcg aaactctaga aatataaaga acatgaaaaa cttttaaatg
     4141 gttttcacac ttgattgcag gtaaaataat ttataccgga gaaaactcca agtgtgaaga
     4201 atataacttt taaccagtgt tcacaactta ttgtacagga aagcatttat agttgagaaa
     4261 tggtgtacaa atataaagaa tgtggaaaag tcattaatat ttgctcacat attacacatc
     4321 agagagttca tacttaataa aagtattata aatacaatta ctgtcaaatg atctttcaga
     4381 aatataaacc tttaaagtga agaagagttc attctaaaga caaacattac aaatataaag
     4441 gggtttgtag tgcttttact tgtatcacag attttattgt acacattttg tactagagaa
     4501 taaccctgaa gcagttgcta aaattttgtt cagcattagg aaatttatgt tggagaagaa
     4561 ttcagcaagt gtgattaatt tggaaaaaca ttttttccat aactatagct tagaaaatgc
     4621 cagagagctc atactaaaat atacttttgc agatgcaata aatacaaaga aatatttaat
     4681 tcaaattgag tttatataat tatcagagaa ttaacagtag aaatagctaa ggcactgaca
     4741 cttcagagac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4801 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4861 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4921 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4981 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5041 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5101 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5161 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5221 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5281 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5341 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5401 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5461 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5521 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5581 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5641 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5701 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5761 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5821 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5881 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5941 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6001 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6061 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6121 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6181 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6241 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6301 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6361 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6421 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6481 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6541 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6601 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6661 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6721 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6781 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6841 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6901 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6961 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     7021 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     7081 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     7141 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     7201 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     7261 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     7321 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
