LOCUS       Exported                7124 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF639 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF639-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7124)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7124)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7124
                     /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             3730..3750
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3763..3807
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3847..3870
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3898..4617
                     /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(3943..3964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4204..4223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4655..5240
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4821..4838)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5048..5067
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5322..6125
                     /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(5385..5404)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5995..6014
                     /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 tcggtaccca attgctgtag aagtgcatgc gatttctgag
     2701 gattatgata tagagacaga aaacaattcc tctgagagtc tccaagacca aactgatgaa
     2761 gaaccgccag ctaaactttg taaaattctt gacaagagcc aagctttgaa tgtgactgcc
     2821 cagcagaaat ggcctttact gagagctaat agcagtggcc tctataaatg tgaactttgt
     2881 gagtttaaca gcaaatattt ttctgactta aagcagcata tgatcctgaa gcataaacgt
     2941 actgattcaa atgtgtgtcg agtatgcaag gaaagtttct ctaccaatat gcttctgata
     3001 gaacatgcca aactgcatga agaggatccc tacatttgta aatactgtga ttataagaca
     3061 gtaatttttg agaacctcag ccagcacatt gcagacaccc attttagtga tcacctctat
     3121 tggtgtgaac agtgtgatgt acagttctcc tcaagcagtg aactctacct acatttccag
     3181 gagcacagct gtgatgaaca gtacttgtgt cagttctgtg aacatgaaac taatgatcca
     3241 gaagacttgc atagccatgt ggtaaatgag catgcatgta aattaataga gttaagtgat
     3301 aagtataaca atggtgaaca tggacagtat agcctcttaa gcaaaattac ctttgacaaa
     3361 tgtaaaaact tctttgtatg tcaagtatgt ggttttcgga gtagacttca cacaaatgtt
     3421 aacaggcatg ttgctattga acatacaaaa atttttcctc atgtttgtga tgactgtggg
     3481 aaaggctttt caagtatgct agaatattgc aagcatttaa attcacattt atctgaaggg
     3541 atttatttat gtcaatattg tgaatattca acaggacaaa ttgaagatct taaaattcat
     3601 ctagatttca agcattcagc tgacttgcct cataaatgta gtgactgctt gatgaggttt
     3661 ggaaatgaaa gggaattaat aagtcacctt ccagtccatg agacaactgg gattccaact
     3721 actgcaagcg agaatcttta ttttcagggc gccgccaaat tcaaagaaac cgctgctgct
     3781 aaattcgaac gccagcacat ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg
     3841 tcgagcctgg aagttctgtt ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg
     3901 gtgagcaaag gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4021 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4081 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4321 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4381 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4501 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctt
     4561 aaggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga
     4621 tctagagtcg agttaattaa gaattccgcc cccccccctc tccctccccc cccctaacgt
     4681 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     4741 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     4801 cattcctagg ggtctttccc ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa
     4861 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     4921 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     4981 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     5041 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     5101 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     5161 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     5221 atgataatat ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag
     5281 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgggatcg gccattgaac
     5341 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5401 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5461 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5521 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5581 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5641 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5701 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5761 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5821 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5881 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5941 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6001 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6061 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6121 tctgagtctg attattctct ttaacttaca gaatgttagt ttaaaataat aaattcatcc
     6181 tttttttgga gatgattaaa tggatgattg taaacacaac ttatgaaatc tgcctttaac
     6241 aagtaacttt tttaaattat aaaattttat tggcattgct ccattttctg tatataaata
     6301 tatctttaat gtggtatttt caattgcgtg atagtttgta gtttcaacca ctcttggtga
     6361 ctgtcatcct gtttcttcca tattctctga tttcatgaat tgaaaagaaa caaatgtatt
     6421 gaagaagtga gctacagttt tccttcctta accatgggtg ctagtaactt tttaaaactc
     6481 aagacaagat tagtttttta tgtgtgaagt cattaaatta ttacacgacc agaactaaaa
     6541 tgcaatatac agttaagtcc acggatactc ccattaatga gaaataacac taggaagcca
     6601 ctattacagg aagaaaagat ttggttttca tggcagtctg tttttttaaa aaaaaatttt
     6661 tgagccacta tctattgttg aatattttaa gatgggatga gggaggaact aataagggct
     6721 tacacaataa aaaataacta tatcataact cattcataac ttgatgtttc attttctgtt
     6781 gaggaaccat aaattcattc acagacttaa tatttttttc ttagagatgg tctcgttctg
     6841 ttgtccagga tggagtgcag tggttgatca tagctcccgg gctgtagtct cccaggctcg
     6901 agcaatcctc ccacttcacc ctcctgcata gctaggacta caggcatgtg gcaccatgcc
     6961 cgctaagttt ttaaatttct tgtagagatg agatctcgct atgttgccca ggccagtctc
     7021 aaactcctgg actcaagcaa tcctcccacc ttggccttcc aaatcactgg gattataggc
     7081 atgagccatt atgcctgact cttgcccaaa tttctgatgt caaa
//
