LOCUS       Exported                6708 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF266 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF266-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6708)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6708
                     /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             3513..3533
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3546..3590
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3630..3653
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3681..4400
                     /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(3726..3747)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3987..4006)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4438..5023
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4604..4621)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4831..4850
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5105..5908
                     /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(5168..5187)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5778..5797
                     /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 ttcttctagt
     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 ctttaaggat tgtgggagag ccttcactgt ttcctcttgc ttaagtcaac
     2701 atatgaaaat ccatgtgggt gagaagcctt atgaatgcaa ggaatgtggg atagccttca
     2761 ctagatcttc tcaacttact gaacatttaa aaactcacac tgcaaaggat ccctttgaat
     2821 gtaagatatg tggaaaatcc tttagaaatt cctcatgcct cagtgatcac tttcgaattc
     2881 acactggaat aaaaccctat aaatgtaagg attgtgggaa agccttcact cagaactcag
     2941 accttactaa gcatgcacga actcacagtg gagagaggcc ctatgaatgt aaggaatgtg
     3001 gaaaggcctt tgccagatcc tctcgcctta gtgaacatac aagaactcac actggagaga
     3061 agccttttga atgtgtcaaa tgtgggaaag cctttgctat ttcttcaaat cttagtggac
     3121 atttgagaat tcacactgga gagaagccct ttgagtgcct ggaatgtggt aaagcattta
     3181 cgcattcctc cagtcttaat aatcacatgc ggacccacag cgccaaaaaa ccattcacgt
     3241 gtatggaatg tggcaaagcc tttaagtttc ccacgtgtgt taaccttcac atgcggatcc
     3301 acactggaga aaaaccctac aaatgtaaac agtgtgggaa atccttcagt tactccaatt
     3361 cgtttcagtt acatgaacga actcacactg gagagaaacc ctatgaatgt aaggagtgcg
     3421 ggaaagcctt cagttcttcc agttcctttc gaaatcatga aagaaggcat gcggatgaga
     3481 gactgtcagc agggattcca actactgcaa gcgagaatct ttattttcag ggcgccgcca
     3541 aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag
     3601 gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct gttccagggg cccctgtcgt
     3661 cgagcggtcc ctcgggttcg atggtgagca aaggcgagga gctgttcacc ggggtggtgc
     3721 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3781 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3841 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3901 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3961 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4021 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4081 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4141 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4201 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4261 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4321 agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc actctcggca
     4381 tggacgagct gtacaagtaa tgatctagag tcgagttaat taagaattcc gccccccccc
     4441 ctctccctcc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     4501 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     4561 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     4621 cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     4681 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     4741 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     4801 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     4861 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     4921 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     4981 gtggttttcc tttgaaaaac acgatgataa tatggccaca acctgttaca ttgcacaaga
     5041 taaaaatata tcatcacgaa cagtaaaact gtctgcttac ataaacagta atacaagggg
     5101 tgttatggga tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5161 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5221 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5281 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5341 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5401 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5461 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5521 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5581 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5641 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5701 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5761 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5821 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5881 ctatcgcctt cttgacgagt tcttctgagg aatgtgggaa aacctaaagg tgtccctgtt
     5941 ctctctgaag acatgaaaac tcactgggga gaaaccctat gaatgtaaaa atgtggaagc
     6001 aactttgtat ctcaggtctt aatgaacaca tatgaattca cagtggagaa gaccctgcat
     6061 cagggaatgt ggaaatgact ttgctgaatt ctcaagcctt accaaacaca tcagaaatct
     6121 cactggagag aaaccgtatg aatgtagaga atctgggaat acctttctga atcccacaaa
     6181 ccttaatgtg tgtatgtgaa ctcacattgg agagaaaccc tgcaatttaa atggtatggt
     6241 ctggatgatg ccccactcca tatttgtaag ccctaagtcc tagttcctta cactataact
     6301 gtatttggac atagggtttt caaacaggtg agtaacttca aatgaggttg ttgggttcga
     6361 tccctaatct gacatcactg gtgtccctat aagggaaact gaaggaagga tacacatgga
     6421 gaagactgtg tggatccacc agaagatggc catctacaag ccaaggacag agacctggaa
     6481 cagatgcttt cattatggcc tccagaggaa accaaccctg tctccacctt gatattgcac
     6541 ttccaggctc cagaactgtg aggcaataaa tttctcttgg ttaaatcatt cagtctgtta
     6601 ttttgtacag caaccctagg aaactaatac tgtgaggaac ttgggaaaag ctttagatca
     6661 agcttgtcca acccgcaggc caggatggct ttgaatgcag accaacac
//
