LOCUS       Exported                6963 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF192 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF192-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6963)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6963
                     /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             3645..3665
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3678..3722
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3762..3785
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3813..4532
                     /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(3858..3879)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4119..4138)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4570..5155
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4736..4753)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4963..4982
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5237..6040
                     /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(5300..5319)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5910..5929
                     /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 agaccaggag tgagaacagg gaattagctt
     2701 caaaacaggt aatatctact ggaatccagc cacatggaga gacagctgcc aaatgcaacg
     2761 gggatgttat caggggtctt gagcatgaag aagcccgaga ccttctgggc agattagaga
     2821 ggcagcgggg aaatcccaca caagagagac gacataaatg tgatgaatgt gggaaaagct
     2881 ttgctcagag ctcaggcctt gttcgccact ggagaatcca cactggggag aaaccctatc
     2941 agtgtaatgt gtgtggtaaa gccttcagtt acaggtcagc ccttctttca catcaggata
     3001 tccacaacaa agtaaaacgc tatcactgta aggagtgtgg caaagccttc agtcagaaca
     3061 caggcctgat tctgcaccag agaatccaca ctggggagaa gccatatcag tgcaatcagt
     3121 gtgggaaggc tttcagtcag agtgcgggcc ttattctgca ccagagaatc cacagtggag
     3181 agagacccta tgaatgtaat gagtgtggga aagctttcag tcatagctca cacctcattg
     3241 gacatcagag aatccacact ggggagaagc cctatgagtg tgatgagtgt gggaaaacct
     3301 tcaggcggag ctcacatctt attggtcatc agaggagcca cactggggag aaaccctaca
     3361 aatgcaatga gtgtgggagg gccttcagtc agaagtcagg ccttattgaa catcagagaa
     3421 tccacactgg agaaagaccc tataaatgta aagaatgtgg gaaagctttc aatgggaaca
     3481 ctggtctcat tcaacacctg agaattcaca caggggagaa gccctaccaa tgtaatgagt
     3541 gtgggaaagc ctttattcag aggtcaagtc tcattcgaca tcagagaatc cacagtggtg
     3601 aaaaatctga atccataagc gttgggattc caactactgc aagcgagaat ctttattttc
     3661 agggcgccgc caaattcaaa gaaaccgctg ctgctaaatt cgaacgccag cacatggaca
     3721 gcggaggtgg aggttcgagc ggtccctcgg gttcgtcgag cctggaagtt ctgttccagg
     3781 ggcccctgtc gtcgagcggt ccctcgggtt cgatggtgag caaaggcgag gagctgttca
     3841 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3901 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3961 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4021 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4081 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     4141 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     4201 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     4261 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     4321 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4381 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     4441 aagaccccaa cgagaagcgc gatcacatgg tccttaagga gttcgtgacc gccgccggga
     4501 tcactctcgg catggacgag ctgtacaagt aatgatctag agtcgagtta attaagaatt
     4561 ccgccccccc ccctctccct ccccccccct aacgttactg gccgaagccg cttggaataa
     4621 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     4681 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     4741 gccaaaggaa tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     4801 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     4861 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     4921 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     4981 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     5041 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     5101 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta
     5161 cattgcacaa gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag
     5221 taatacaagg ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc
     5281 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5341 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5401 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5461 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5521 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5581 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5641 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5701 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5761 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5821 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5881 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5941 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6001 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gtctaggaac aacatcagtt
     6061 agagtttgag cattattcag cattagggaa accacacact ggtgagaggt ctttcagtgt
     6121 actaaaaggc agaaaggtca tcacaacttt agtgtcagaa tctatagtgg tggcaaagtt
     6181 aggatagctc tttagtaatt tgggcccagt gccttggtga aggttgatta gcttgactgt
     6241 ctttgaaagt atctgatgga gctcctgatg acctaatgta tcctttagaa atttaaaata
     6301 gcattagagc aagttgcttg tcatggcttg aagagcttaa ctttgtcttt tgggtgagta
     6361 gctataggtt tgagagaggc tggctactcc tagttcctgt gcttcttccc atctcctgtg
     6421 atccccctct cccatttatt cccttgaagg tgcccttgta ttcctaatct gatctaagaa
     6481 gctgctgtag agtcagaagt agcttctgtg tgaaacttat atttgtatgt agctttctag
     6541 gaaaatttta cagacactta aagtatttat gcttaagtgt gcattttttt attctaatat
     6601 tccttctagt cagtggaaat ttgtattccc atacaaactc agaatgctat atttttaaca
     6661 gcactccagc atttttcttc atgtatcacc ttcactgact ccattgagtc attgagcatc
     6721 tgtacatatc cttcaccacc cccagtccca gcacctgtgt cagtaaagaa agtggacaca
     6781 ttagtaatgg ttatgggagt aatacagaga aagaggtgag tggagactca gcccagctgc
     6841 tcttgagctt ggatagttat acattttggt gacttctgac tctccttcca cctccccact
     6901 gtacactgtg gtacagatag ttggcactat cctagttatg gcatcaggta ggagaaggag
     6961 acg
//
