LOCUS       Exported                7341 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF423 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZNF423-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7341)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7341
                     /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             3880..3900
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3913..3957
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3997..4020
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4048..4767
                     /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(4093..4114)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4354..4373)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4805..5390
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4971..4988)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5198..5217
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5472..6275
                     /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(5535..5554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6145..6164
                     /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 tagaaggaca 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 tcggtaccct ccgggcgagg ctactggcat cacctggtcg
     2701 ccctcgccgc ggacccgagc ccaggagaga cgcggtgcct ctgctgaccc ctggcggtca
     2761 gcggcacagc tgcaggagtc gccgggccct ggggggcgcc cagaaggagc ctctcccggg
     2821 gatctagggt cctgtgcggc cccaaacagg gccaccctac acctggtgga tgttgcctcc
     2881 tttctccagg tgtcatgagg tggagcacca cctgaggctt tgatgaaatg ggtctgaatt
     2941 tggaacacga accactgaaa agaaacaaaa agacggcttt cagatgccaa atcccacccc
     3001 cagtcccagg ccgctggctc ctgtgaaaca gatcaaccct gctggccccc tctggctggc
     3061 agggggagga aatgccattg actgtaatga gagggaaaca tttggccaca gggtcccacc
     3121 tgggccctgg attcctaggt gcattcggcc aaactgcact gagtccctgt ctctggttgg
     3181 gccattgcca tgtgaccaag caggcaggct cccggagccc agcagcagag gctcctcccg
     3241 cgtcttcctg ccgggtgacc tgtcccctgc ctgggccgca ggcaccacgg ccgcccactg
     3301 cctcagcacg cctgtgcccg ctcgggtgtt tcttggccgg agagctgcac agcacatctg
     3361 ctgtctccct ctgctaggct cggagcagcc acggggaaga ctgagagggg gatgcgaggg
     3421 cagcctgcag gcgatcagag tttcctcctc ttcattagag gaaaagggag atggagaggt
     3481 gaacagggat gtttttaatt cctgcctcct ccccggcatt tcccctttcc ctccacctag
     3541 ccagtccccc cggtaagacc tgaatggtct ttaaaaaaaa aaaaaaaaaa aaagccaaaa
     3601 aacaacatat agggagtcca agttggcagt aacaggaacc tatccccaag ccccaaatca
     3661 acgtggcgtc ttggcagctg cgtccctgga gcactttttc cttgttgcat tttgttttgt
     3721 tataatcgag acgcttttct gcggctcccg gcgtaaatgc actgagtggg agggacgagc
     3781 atgctctcca ttcttctcct tcatgtgtgt ctcctttctt cgcctttgca gaaccacacg
     3841 atgagccagc acgcacaggg gattccaact actgcaagcg agaatcttta ttttcagggc
     3901 gccgccaaat tcaaagaaac cgctgctgct aaattcgaac gccagcacat ggacagcgga
     3961 ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt ccaggggccc
     4021 ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag gcgaggagct gttcaccggg
     4081 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4141 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4201 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     4261 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4321 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     4381 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     4441 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     4501 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     4561 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4621 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     4681 cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc cgggatcact
     4741 ctcggcatgg acgagctgta caagtaatga tctagagtcg agttaattaa gaattccgcc
     4801 cccccccctc tccctccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     4861 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     4921 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     4981 aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     5041 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     5101 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     5161 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     5221 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     5281 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     5341 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc tgttacattg
     5401 cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata aacagtaata
     5461 caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt tctccggccg
     5521 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5581 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5641 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5701 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5761 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5821 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5881 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5941 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6001 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6061 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6121 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6181 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6241 tcgccttcta tcgccttctt gacgagttct tctgagtcgg gatcgctcaa caggacacct
     6301 ctccgcagaa ggcttgccgg agacgccgtg gggagggcca tttgaacatt acatccaatc
     6361 aaagtgtcat ttgcaaccca gatgtaaaac tctaatgatt tggccatgag gcgctgctat
     6421 tataagcagc tggaaatgaa tattaatggc agagattaaa agtattccat gctcagtatt
     6481 ttttattgtc ctgctacagc tagtgtgctt ttagactttc cgccgcagac tacatttcta
     6541 gagttagaga aacctgcttt ttaaggctat tgtcctttgt tccttcatgt attatattga
     6601 tagtttttaa aaaagaatta gtgtgatttt ttttctttgc ttcttttttt tctttcttgt
     6661 ttttcttccc ccccaccccc cacccccttc ggttaactac tttttaattg caattctagg
     6721 taattgtgca tcgtgatgtg attgcttggc tattgtctga atatttcctt ttaatttttt
     6781 aattaaagac taatgctttg attggatttg ccagttcacc ggacagtgat taaaactatg
     6841 taatgaatat aatcggtttc agtgcaactg gatggtctgc ttttaaatgt gacttaatct
     6901 gactgcagta actagtacag ttcaataaag ggaatccatg cgattagcat ccggctgcct
     6961 ggttctctcc tccccagcct agggccgcgt ggggggctgt cagctcctgt aaggtgtgtt
     7021 cccctacaag ttcctgtggg cttgcacacg cccttgggga cctgcacaca atagaccctg
     7081 ctccccttgg aaaagaatcg tagtaaccta atgcatgcag actggacagg ctggcagtgg
     7141 ccacaggagt caaagcagaa gaccccttaa tgatggaata ccctgatctt cccactcttg
     7201 gatggggaaa cagaggccca agggtaacgt gcagctttcc caaggccacg cagcaggttg
     7261 gtcaccatgc tgattgtgaa gtcttcttgg ctttcccttc agggttggct ttcccctttg
     7321 ttggatacag gcaggcaagt g
//
