LOCUS       Exported                7138 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZBTB9 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZBTB9-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7138)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7138
                     /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             3890..3910
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3923..3967
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4007..4030
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4058..4777
                     /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(4103..4124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4364..4383)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4815..5400
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4981..4998)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5208..5227
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5482..6285
                     /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(5545..5564)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6155..6174
                     /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 tgacaagctg cttctggggg atgcgcctcg
     2701 tctcactcta ccgagtgtca ttgaagccga tgccttcgag gggctgctcc agctcattta
     2761 ttcagggcgt ctccgcctgc cactggatgc tcttcctgct catctccttg tggccagtgg
     2821 ccttcaaatg tggcaggtag tagatcagtg ctcagaaatt cttagagaat tagaaacttc
     2881 aggtggtgga atttcagccc gtggaggaaa ctcctaccat gcccttcttt ccactacatc
     2941 ctctacagga ggctggtgca ttcgctcttc gcctttccag accccagtac agtcctctgc
     3001 ttctactgaa agccctgctt ccactgagag ccctgtggga ggggagggaa gtgaactggg
     3061 agaagtgctg caaattcagg tggaagaaga agaggaggag gaggaagatg atgatgatga
     3121 ggaccagggg tcagccacac tctctcagac tcctcagccc cagagagtat caggggtttt
     3181 tccccgtcct catggacccc acccactgcc catgactgct actccccgaa agcttccaga
     3241 gggtgagagt gcaccacttg agcttcctgc ccctcctgca ctgcccccca aaatcttcta
     3301 cattaagcag gaacccttcg agcctaagga ggagatatca ggaagcggaa ctcagcctgg
     3361 aggagcaaag gaggaaacca aagtgttttc tggaggggac actgaaggga atggggagct
     3421 agggttcttg ttgccttcag ggccagggcc aacatctggg ggagggggtc catcctggaa
     3481 accagtggat cttcatggga atgaaatcct gtcagggggt ggaggacctg ggggagcagg
     3541 ccaggccgtg catgggcctg tgaagctagg ggggacaccc cctgcagatg gaaaacgctt
     3601 tggttgcctg tgtgggaagc ggtttgcagt gaagccaaag cgtgaccggc acatcatgct
     3661 gaccttcagc cttcggcctt ttggctgtgg catctgcaac aagcgcttca agctgaagca
     3721 ccatctgaca gagcacatga agacccatgc tggagccctg catgcctgtc cccactgtgg
     3781 ccgtcggttc cgagtccatg cctgttttct ccgccaccgg gacctatgca agggccaggg
     3841 ctgggccact gcccactgga cttacaaggg gattccaact actgcaagcg agaatcttta
     3901 ttttcagggc gccgccaaat tcaaagaaac cgctgctgct aaattcgaac gccagcacat
     3961 ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt
     4021 ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag gcgaggagct
     4081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4141 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     4201 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     4261 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4321 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     4381 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4441 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4501 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     4561 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4621 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     4681 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc
     4741 cgggatcact ctcggcatgg acgagctgta caagtaatga tctagagtcg agttaattaa
     4801 gaattccgcc cccccccctc tccctccccc cccctaacgt tactggccga agccgcttgg
     4861 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     4921 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     4981 ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     5041 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     5101 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     5161 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     5221 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     5281 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     5341 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     5401 tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata
     5461 aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt
     5521 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5581 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5641 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5701 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5761 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5821 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5881 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5941 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6001 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6061 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6121 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6181 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6241 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtcct gctgaggcta
     6301 tacactagct tctagaacaa gataaccact gctgctgatg gatacttttc cctcactgcc
     6361 atggcacacc agtcatggat cttgtaatca tgccaagaga atagatacat tatggacctc
     6421 ttgttcttag atatgggcct ctcagcctgg cagatgtgga aactcaaatt tctcgtccca
     6481 ctccaggttt tggctagcca accctgcagg aaagtggttt ataggccatt catacttaag
     6541 ttgatcactt gcccatggtg gacatttttg tggtggtgat gtccattaag gaaaccagat
     6601 tttcaattat ttagtgagag aagagttaga gcaaaagaca gtggtaaatg ttttattccg
     6661 tctccatgag gaattgaagg agttggtctc cacctagaga tacatttgat ttacagctta
     6721 agtaattcag aggctaagct ctaagctttt ttctctcatt gctggaatga tttaagcaga
     6781 agtccttttg tgtactttta aaattgtatc tttccaggag cccctcagat tgtaccttgc
     6841 tttctcacca atagacacct tcccgacact tttttaatgt tgtagctgag cactttaaca
     6901 agttgagcat tccatgtttc attcttagaa ccttctttaa tagagggtct tccctcaaca
     6961 gcctgtgcct ctggtctacc tttgaccacc actgataact aatatattgg tcacaatgac
     7021 tggaatgtga ctagtgatct caggagatgg cactgtccta aagtgctgtc agggtggcac
     7081 cactgctctc tgaacaactt accttggtca gagggactca ggtttgggac agcacaag
//
