LOCUS       Exported                6769 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF696 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF696-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6769)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6769)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6769
                     /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             3528..3548
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3561..3605
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3645..3668
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3696..4415
                     /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(3741..3762)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4002..4021)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4453..5038
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4619..4636)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4846..4865
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5120..5923
                     /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(5183..5202)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5793..5812
                     /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 cttggtcact tctgagaaaa ctggaggaca gagtggcctc gagtcagacg
     2701 tccctccgaa cgcaggcccc ggcgcagagg gcgggggcag ctggaagggg cggcctttcc
     2761 cgtgcggcgc ctgtggccgc agcttcaagt gctcctcgga cgcggcaaag caccggagca
     2821 tccactcggg ggagaaaccg tacgagtgca gcgactgcgg gaaggccttc atccacagct
     2881 cgcacgtggt ccggcaccag cgggcgcaca gcggggagag gccctacgcg tgcgccgagt
     2941 gcggcaaggc cttcggccag agcttcaacc tcctccggca ccagcgcgtg cacacgggcg
     3001 agaagcccta cgcgtgcgcc gactgcggca aggccttcgg ccagaggtcg gacgccgcca
     3061 agcaccgccg cacccacacc ggggagaggc tgtacgcgtg cggcgagtgc gggaagcgct
     3121 tcctgcacag ctcgaacgtg gtccggcacc ggcggaccca ccacggggag aacccgtacg
     3181 agtgccggga gtgcggccag gccttcagcc agagctccaa cctcctccag caccagcgcg
     3241 tgcacacggg ggagcggccc ttcgcctgcc aggactgcgg ccgcgccttc agccgcagct
     3301 ccttcctccg cgagcaccgc cgcatccaca ccggggagaa gccccaccag tgcggccact
     3361 gcgggcgcgc gttccgggcg ctgtcgggct tcttccggca ccagcgactc cacacgggcg
     3421 agaagccgtt ccgctgcacc gagtgcggcc gcgccttccg cctgagcttc cacctcatcc
     3481 agcaccggcg ggtgcatggc gccgagggga ttccaactac tgcaagcgag aatctttatt
     3541 ttcagggcgc cgccaaattc aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg
     3601 acagcggagg tggaggttcg agcggtccct cgggttcgtc gagcctggaa gttctgttcc
     3661 aggggcccct gtcgtcgagc ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt
     3721 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3781 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     3841 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     3901 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3961 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     4021 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     4081 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     4141 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     4201 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     4261 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     4321 gcaaagaccc caacgagaag cgcgatcaca tggtccttaa ggagttcgtg accgccgccg
     4381 ggatcactct cggcatggac gagctgtaca agtaatgatc tagagtcgag ttaattaaga
     4441 attccgcccc cccccctctc cctccccccc cctaacgtta ctggccgaag ccgcttggaa
     4501 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4561 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4621 ctcgccaaag gaatgcaggg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     4681 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4741 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     4801 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     4861 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     4921 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     4981 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacctg
     5041 ttacattgca caagataaaa atatatcatc acgaacagta aaactgtctg cttacataaa
     5101 cagtaataca aggggtgtta tgggatcggc cattgaacaa gatggattgc acgcaggttc
     5161 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5221 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5281 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5341 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5401 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5461 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5521 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5581 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5641 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5701 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5761 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5821 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5881 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagccgggg ctgcggcgga
     5941 agagatgccg gcggcctggt gggcgcgagg ccgaggccgg gggaggctcc tgtccgcccc
     6001 gtgcgcggtg aggaagtaac agccggctgc gcgcctggtt ctcgggttgc gaaagcctcg
     6061 ccaggcctgc atccgccttg gcttgggagc aatcaggaga gacagggctc ggggaaggcg
     6121 agcgctgccc gcgggaggcg attcccagag gcggggaggt ctcaggggtc tgtcccgggc
     6181 cggccgcccg cctctgagac tccccgcctc ccacggtcag ctgctgcggg gcagtggccg
     6241 gtgattgaat cggtcattcc tccgcaacca ggcctccatc aaaaccctgc atgagatgag
     6301 ggcctcgggc ttcgggttgg tgaaggcggg gtgggggggc agttaggaag ggggcgccca
     6361 gacagggcat ggaagcctcg cgtcctcccc cgtaccttgc gggtcggtgg ttcatccatg
     6421 tatttcaacg ccttttcaca gccggttcat atggtcccga cttctgtgtg cttagaaaat
     6481 tatggaactt ggccgggtgc ggtggctccc gcctgtaatc ctagcacttt gggaggccga
     6541 ggcgggcgga tcacacggtc aagagatcaa gaccatcctg gccaacatgg caaaaccccc
     6601 atctcgacta aaaatacaaa aaattaattg ggcgtggtgg cgtgcgcctg taatctcagc
     6661 tacttgggag cctggggcag gagaatcact tgaacctggg aggcggaggt tgcagtgagc
     6721 ggagatctgg gcactgtact ttagcctggg cgacagagag acccatctc
//
