LOCUS       Exported                7108 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF331 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF331-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7108)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7108
                     /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             3741..3761
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3774..3818
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3858..3881
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3909..4628
                     /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(3954..3975)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4215..4234)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4666..5251
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4832..4849)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5059..5078
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5333..6136
                     /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(5396..5415)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6006..6025
                     /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 tcggtaccca gagaaggcac ccctcctaga acacatcaga
     2701 gacatcataa ggagaattcc tttgaatgta aggactgtgg gaaggccttt agtcgtggct
     2761 atcaacttag tcaacatcag aaaatccata ctggtgagaa accttatgaa tgtaaagaat
     2821 gtaagaaggc cttccgttgg ggcaatcagc ttactcaaca tcaaaaaatt catactgggg
     2881 agaagcccta cgaatgtaaa gactgtggga aggcttttcg atggggctca agcctcgtta
     2941 ttcataagag gattcatact ggtgaaaaac cctatgaatg taaagactgt ggaaaggcct
     3001 ttcggcgtgg tgatgagctc actcagcacc agagattcca cactggggag aaagactacg
     3061 aatgcaaaga ctgtgggaag acctttagcc gtgtgtataa acttattcag cacaagagaa
     3121 ttcatagtgg ggagaagcct tacgagtgta aagactgtgg gaaggctttt atttgtggtt
     3181 caagcctcat tcagcataaa agaattcaca caggtgagaa accctatgaa tgtcaagaat
     3241 gtgggaaggc ctttactcga gtcaattacc ttactcagca tcagaagatc cacaccggtg
     3301 agaagcctca cgaatgtaag gagtgtggga aggcctttcg ctggggttcg agcctcgtta
     3361 agcacgagag gatacatacg ggcgagaagc cgtacaagtg cacagaatgt gggaaggcct
     3421 tcaattgtgg ctatcacctc actcagcacg agagaatcca cacaggcgaa accccgtata
     3481 aatgtaagga gtgtgggaag gctttcattt atggatcgag cctcgtgaaa catgagagaa
     3541 ttcataccgg ggtgaaaccc tatgggtgta cagaatgtgg gaagagcttt agtcacggcc
     3601 atcagcttac acaacatcag aaaacgcaca gtggggcgaa atcctacgaa tgtaaggagt
     3661 gcgggaaggc atgtaaccac ctaaaccatc tccgagaaca tcagaggatc cacaacagtg
     3721 ggattccaac tactgcaagc gagaatcttt attttcaggg cgccgccaaa ttcaaagaaa
     3781 ccgctgctgc taaattcgaa cgccagcaca tggacagcgg aggtggaggt tcgagcggtc
     3841 cctcgggttc gtcgagcctg gaagttctgt tccaggggcc cctgtcgtcg agcggtccct
     3901 cgggttcgat ggtgagcaaa ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3961 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4021 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4081 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4141 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4201 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4261 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4321 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4381 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4441 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4501 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4561 acatggtcct taaggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4621 acaagtaatg atctagagtc gagttaatta agaattccgc ccccccccct ctccctcccc
     4681 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     4741 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     4801 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca gggtctgttg
     4861 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     4921 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     4981 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     5041 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     5101 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     5161 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     5221 tgaaaaacac gatgataata tggccacaac ctgttacatt gcacaagata aaaatatatc
     5281 atcacgaaca gtaaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgggatc
     5341 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5401 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5461 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5521 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5581 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5641 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5701 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5761 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5821 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5881 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5941 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6001 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6061 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6121 tgacgagttc ttctgagtct gaagagcctt ttgaacgcag tagcccgctc gtatctatgg
     6181 tttcgctttc cacagtttgt tacctgcagt caactgcagt tcaaaaatat taaatggaaa
     6241 attccagaaa taaagaattt taagtctcaa atggtgtgcc cttctgagta gcgtgatgaa
     6301 atctctcgct gtccggctcc agccggccgg ggatgtgagt catcccttgg tccagcacat
     6361 ccacgctgta tacgccaccc accctgctag tgacttagta gccgtcttgg tgatcagatc
     6421 aactatccca gcatcacagt gcctgtgccc aagcagtcct cactttgctt aacagtggcc
     6481 ccagagagca ggagtagtga tgctggtgat tcggatatgc caaagagaag ccacaaagtg
     6541 cttcctttta aatgaaaagg tgaaagttct caacttaaaa aagaaaagaa aaaaatcata
     6601 tactgaggtt gctaagatct acatgacaat aagatatttt gggagagaga tacgttcaca
     6661 taattttatt acatatattg ttacaattgt tctattttgt tagttactgt tggtaatctc
     6721 ttactgtgcc taatttataa attaaacttt agatatgtat gtacaggaaa aaatgtagta
     6781 tatagtatcg tatatatgta caggaaaaaa acgtactatc tgtggtttca ggtatccact
     6841 gcgggtcttc gaacgtaccc ccctcaggtg agaggggact gctgtacaaa gaatatagga
     6901 aggcctctag actacatcca ttccttcctc aacaggagga aaattcctgg tacagtttaa
     6961 cgtaaggctt cactgatcac tgttgttttt gtttttgttt ttgttttttt gagacagagt
     7021 ctcactcttg cccaggctgg agtgcagtgg cacggtcttg gctcactgca acctccacct
     7081 cccaggttcc agcaattctc ctgcctca
//
