LOCUS       Exported                6667 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor BCL6 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pBCL6-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6667)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6667
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(19..36)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(19..35)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(28..50)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(244..263)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     363..385
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(423..441)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        509..613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             614..1474
                     /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     complement(832..851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1645..2233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2134..2153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2387..2404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2521..2542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2557..2587
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2595..2611
                     /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)."
     primer_bind     2600..2622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2619..2635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2619..2635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3540..3560
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3573..3617
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3657..3680
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3708..4427
                     /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(3753..3774)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4014..4033)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4465..5050
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4631..4648)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4858..4877
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5132..5935
                     /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(5195..5214)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5805..5824
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
       61 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      121 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
      181 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
      241 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
      301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
      421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2401 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2461 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2521 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2581 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2641 ttacgccaag cttaagtaac ttttatctaa actaaagcgc ttattaccca ttcattgaat
     2701 taaatcaaga aacggctcca cttgctaatg tttagttagc ttatctagga tatataccct
     2761 aggaaagaat ataactgcat ttaaaaaaat ttaccctaga ctccagaatt tccgtgaatt
     2821 cagactgatc tagtccattt gtccagatgt gttcagatta attgcataat ttgaataaga
     2881 ggtatattct gtgaggtcct aggaagatta taaagtttag gaaaaaaata gccactgccc
     2941 tcaaagtgtt acccactgaa gaaggatcca tgcacacgga gggctgtgtg gtaaaggtag
     3001 aaatcttaca gaggctcaaa ggaaaacaag gattattttc atccttgaga gattcagaaa
     3061 tgccttaatc attagaagag tttgcatcta actgcacttt tggataagac tcaaacttgt
     3121 ggagttaagg cagggaagga cgtctctagc aagggagagg cagaagcaac acttgggaaa
     3181 gcagggaacg gacatttgca caacgctagc ttatctgtcg gcagaagccc ggatgcatat
     3241 gggttcttca cctggccttg gagaggcaag agctgcccca gtccctcgta gggagaatct
     3301 tcctctctgg gtctttgagc aatgacatct ttcttgggtt gatgacagct aacagtttgc
     3361 ctctgatatc cctgcagtgt gagaagtgta acctgcattt ccgtcacaaa agccagctgc
     3421 gacttcactt gcgccagaag catggcgcca tcaccaacac caaggtgcaa taccgcgtgt
     3481 cagccactga cctgcctccg gagctcccca aagcctgcgg gattccaact actgcaagcg
     3541 agaatcttta ttttcagggc gccgccaaat tcaaagaaac cgctgctgct aaattcgaac
     3601 gccagcacat ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg
     3661 aagttctgtt ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag
     3721 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3781 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3841 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3901 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     3961 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4021 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4081 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4141 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     4201 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     4261 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     4321 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg
     4381 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga tctagagtcg
     4441 agttaattaa gaattccgcc cccccccctc tccctccccc cccctaacgt tactggccga
     4501 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     4561 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     4621 ggtctttccc ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt
     4681 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     4741 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     4801 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     4861 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     4921 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     4981 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     5041 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
     5101 tgcttacata aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt
     5161 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5221 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5281 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5341 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5401 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5461 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5521 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5581 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5641 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5701 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5761 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5821 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5881 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaagcat
     5941 ggagtgttga tgctttcgtc tccagcccct tctcagaatc tacccaaagg atactgtaac
     6001 actttacaat gttcatccca tgatgtagtg cctctttcat ccactagtgc aaatcatagc
     6061 tgggggttgg gggtggtggg ggtcggggcc tgggggactg ggagccgcag cagctccccc
     6121 tcccccactg ccataaaaca ttaagaaaat catattgctt cttctcctat gtgtaaggtg
     6181 aaccatgtca gcaaaaagca aaatcatttt atatgtcaaa gcaggggagt atgcaaaagt
     6241 tctgacttga ctttagtctg caaaatgagg aatgtatatg ttttgtggga acagatgttt
     6301 cttttgtatg taaatgtgca ttcttttaaa agacaagact tcagtatgtt gtcaaagaga
     6361 gggctttaat ttttttaacc aaaggtgaag gaatatatgg cagagttgta aatatataaa
     6421 tatatatata tataaaataa atatatataa acctaaaaaa gatatattaa aaatataaaa
     6481 ctgcgttaaa ggctcgattt tgtatctgca ggcagacacg gatctgagaa tctttattga
     6541 gaaagagcac ttaagagaat attttaagta ttgcatctgt ataagtaaga aaatattttg
     6601 tctaaaatgc ctcagtgtat ttgtattttt ttgcaagtga aggtttacaa tttacaaagt
     6661 gtgtatt
//
