LOCUS       Exported                7195 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF764 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF764-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7195)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7195)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7195
                     /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             3878..3898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3911..3955
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3995..4018
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4046..4765
                     /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(4091..4112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4352..4371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4803..5388
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4969..4986)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5196..5215
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5470..6273
                     /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(5533..5552)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6143..6162
                     /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 atcctcaggt gtccgttcct ccttctggat
     2701 caggcccaga tgttcatgtc atctctgccc ttcccccacc gtgacatttc cccagccccg
     2761 aggcggaatg ggctgttctc tccctcccgg ctctgtctgg ctctgcagcc gaggaacagg
     2821 tgtcactcct ccatctccca ggagcagccg gggatccagc aggcctgtcc ctgtccacgc
     2881 gggctgtggg attcaaggac caggatgagc cgagcctgaa ctctcccctc tttatctctc
     2941 agcagattcc agaaacaaga aaaaggaaag acaaagggaa gggacgggag ccctggagaa
     3001 gcccgaccct gtggccgccg ggtctcctgg gctgaagtct ccccaagccc cctcggccgg
     3061 gcccccttat ggttgggagc agctgtccaa ggcaccgcac cggggacgcc cctccctctg
     3121 tgcccacccc cctgtccccc gagctgacca gcgtcatggc tgctacgtgt gcgggaagag
     3181 ctttgcctgg cgctccacac tggtggagca cgtctacagt cacactggcg agaagccctt
     3241 ccactgcact gactgcggca agggcttcgg ccacgcttcc tccctgagca aacaccgggc
     3301 catccatcgt ggggagcggc cccaccgctg tctggagtgt ggccgggcct tcacgcagcg
     3361 ctcggcgctg acttcgcacc tgcgcgtcca caccggcgag aaaccctatg gctgcgccga
     3421 ctgtggccgc cgcttcagcc agagctctgc cctctaccag caccggcgcg tgcacagcgg
     3481 cgagaccccc ttcccctgcc cggactgtgg ccgcgccttc gcctacccct cggacctgcg
     3541 gcgccacgtg cgcacccaca ccggcgagaa gccctacccg tgcccggact gcgggcgctg
     3601 cttccgccag agctcggaga tggcagccca caggcgcacc cacagcggcg agaagcccta
     3661 cccctgcccg cagtgcggcc gccgctttgg ccagaagtca gccgtggcca aacaccagtg
     3721 ggttcatcgg cccggggccg ggggccacag gggccgggtc gccgggcgtc tgtctgtgac
     3781 cctgacccct ggccacggag acctggaccc gcccgtgggc ttccagctgt acccggagat
     3841 attccaggag tgtgggggga ttccaactac tgcaagcgag aatctttatt ttcagggcgc
     3901 cgccaaattc aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg acagcggagg
     3961 tggaggttcg agcggtccct cgggttcgtc gagcctggaa gttctgttcc aggggcccct
     4021 gtcgtcgagc ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt tcaccggggt
     4081 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4141 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     4201 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     4261 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4321 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4381 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4441 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4501 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     4561 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4621 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4681 caacgagaag cgcgatcaca tggtccttaa ggagttcgtg accgccgccg ggatcactct
     4741 cggcatggac gagctgtaca agtaatgatc tagagtcgag ttaattaaga attccgcccc
     4801 cccccctctc cctccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     4861 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     4921 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     4981 gaatgcaggg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     5041 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     5101 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     5161 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     5221 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     5281 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     5341 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacctg ttacattgca
     5401 caagataaaa atatatcatc acgaacagta aaactgtctg cttacataaa cagtaataca
     5461 aggggtgtta tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct
     5521 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5581 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5641 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5701 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5761 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5821 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5881 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5941 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6001 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6061 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6121 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6181 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6241 gccttctatc gccttcttga cgagttcttc tgagtccggc cttaaaagtg actatctaga
     6301 cattacggga ggcccgagat gggctcaggg gcccgaacct ctgcggcggc ctgcagggag
     6361 gtcccagaat ctagggcaag agctggccct ggggtgcgga cagtctgatc ttgggccctt
     6421 agcagcctcc tctgccagca ccttgctccc cactgcccag ggctctccaa ggccccctct
     6481 actgaggcag ggctgagttg agaaccccca gacctacata aggggaacca aaagctgttt
     6541 ctcagcccag aggtgctaag aggactgagg tagaggagaa ccttgtttta cccattgtct
     6601 ttttaaattt tttatcatta tttttattct tgtgaccctc ccagaacaca atactcattg
     6661 tctttttatt tatttattta tttatttctg agacagcttc ctgctctgtc acccaggctg
     6721 gagtgcagtg gtgtgaacag ggctaactgc agcctcaacc tgcagtgctc aaggaatcct
     6781 ctcccatcag ccttccaaat atctgagact acaggcgggc aacactacgg ccagctaatt
     6841 tttaaatttt acatagagat gaggttctca ctgttgtcca ggctggtctc aaactccttg
     6901 gctgaagtga tcctgtcgtc tcggcttccg caagtgctgg gattacaggc gtgagctacc
     6961 accatgcctg gcttccattg tctttgtgtg tgtgtgtgtg tgatggggtt ttgctcttgt
     7021 tgcccaggct ggagtgcaat agcgcaatct cagctcactg caacctctgc ctccagagtt
     7081 caagcaattc tccttcctca gcctcccaag tagctgggac tacaggcacc caccacctcc
     7141 attgtctttt tactgcccaa gttaaaatcc tgtgactgtg tactggatct ggcct
//
