LOCUS       Exported                7287 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor TFCP2 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pTFCP2-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7287)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7287
                     /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             3887..3907
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3920..3964
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4004..4027
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4055..4774
                     /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(4100..4121)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4361..4380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4812..5397
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4978..4995)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5205..5224
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5479..6282
                     /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(5542..5561)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6152..6171
                     /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 gggagaaggg agtttgagaa gggaaagtgg
     2701 ggaaggaaaa ttaaagaggt tttaggaaag agcacatatg ggagttgaag acacggaagt
     2761 cagttctcct gtatgcacat atttgtgtcc gtgtgctcct tggagagtct tggggttaac
     2821 ccagggaaat gcataccagg ctttaaaaca actcttccaa tctgcttagt tttttaatat
     2881 ttaaaaacca aatgggaggg gaggaaaaaa cttttatgat cctagatatt caagaaatac
     2941 attatttgtg acctgtaaag cctcccatat aaatccctct ggatgtgggg tagtccttag
     3001 ggacacagga atataaaggg gttgtttttc atttaaataa tcacaaattt tcttgcattt
     3061 ttcagatgat acagaacttt caggaagaag catgttttat tctggacaca atgaaaggta
     3121 atagtaagac aaacatcttt ctctaagcag tttaacagct ttactactac taggcaaagt
     3181 acatacttct ttgaggagag ataggaaaag taaccatatt tgggtaaaga aagtgacaat
     3241 attagagatt gttccttttt ttaagttggc tttttttttt tttttttttt ttgaaacaga
     3301 gtctcgatct gttgcccagg ctggagtgca gtggcatgat ctcggctcac tacaaccttc
     3361 gcttcccaag ttcaagtgat tctcctccct ctgcctccca agtagctggg attacaggca
     3421 cgtgccacca tgtctggcta atttttgtat ttttagtaga gatgggtttc accttgttgg
     3481 ccaggctggt ctcgaactcc tgacctcaag cgatccaccc accttggcct cccaaagtgc
     3541 tgggattaca gacatgagcc actgtgcctg gcccagcttt ttgccattct cttccatctt
     3601 ccattacaga tgaggaaatt gaggcatgtt gtttttataa tgcatttttt ttgagacaga
     3661 atttcgctct tgttgcccaa gctggagtcc agtggcgtga tctcagctca ctgcaacctc
     3721 cacctccctt ttctcccctt ctcacgagaa tcttaccata ctttttttct cctgctgtgt
     3781 tttagaggtg actaaagatg attctctctc tctctttttt ttaacagcag aaaccaatga
     3841 tagctatcat atcatactga agcccgggat tccaactact gcaagcgaga atctttattt
     3901 tcagggcgcc gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga
     3961 cagcggaggt ggaggttcga gcggtccctc gggttcgtcg agcctggaag ttctgttcca
     4021 ggggcccctg tcgtcgagcg gtccctcggg ttcgatggtg agcaaaggcg aggagctgtt
     4081 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4141 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4201 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4261 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4321 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4381 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4441 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4501 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4561 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4621 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4681 caaagacccc aacgagaagc gcgatcacat ggtccttaag gagttcgtga ccgccgccgg
     4741 gatcactctc ggcatggacg agctgtacaa gtaatgatct agagtcgagt taattaagaa
     4801 ttccgccccc ccccctctcc ctcccccccc ctaacgttac tggccgaagc cgcttggaat
     4861 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4921 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4981 tcgccaaagg aatgcagggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     5041 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5101 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5161 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5221 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     5281 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     5341 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaacctgt
     5401 tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc ttacataaac
     5461 agtaatacaa ggggtgttat gggatcggcc attgaacaag atggattgca cgcaggttct
     5521 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5581 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5641 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5701 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5761 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5821 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5881 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5941 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6001 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6061 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6121 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6181 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6241 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagagtgcgg cgtttcgtgc
     6301 ccagtggctg ctccttcctt cacctctgaa aacggccctc ttgaaggggg atatgaatgg
     6361 agatttgaag gtctgcaaga acctgactcg tctgactgtg tgtggaggag tccaggccat
     6421 ggaggcagaa tcctggccct ctgtgttggc ccaagctctt gtggtacaca cagattactg
     6481 cccaatatgc agttctgcag ctgttttagt taaatttctg gaccttgttg ttgttaaata
     6541 tcagtagaaa ctctacataa tttagagtgt atgtagggca taatgatgat gggaattgtg
     6601 tgatgtttaa caggaagatc ttaaattttg tgatatggag ccctgtaatt tttttcttat
     6661 ataaaaatgg gtatctatat tcataagact aggtcttcaa tctctttgta ctggtctcaa
     6721 atgtactggt atctctgctt tttgccacag tttgtccctg aaaactttat cagtgaggag
     6781 aaatacagaa ttttcctttt ggttcccctg taataccaca actaatagta ttttcagcta
     6841 acatttattg accaggcagt gtgataaatt ttttgaatga ccatcttgtt taacccccta
     6901 atcacactat ctgaagtagg tgctatagtg acccccaacc gaagatgagg aaatggagac
     6961 acacagtggc taagtagctt gctaggtgcc aggagctggc aggcagtgat gctgaaatcc
     7021 aaaccaggca atctggccct ccttattcac cctctcatac caccagtgct acccattata
     7081 acctgtgttc acttcgctta ttactgtggc ccttctggtg actacaacat aacactgagt
     7141 atgaactgag gaattaacca ctgaaagaga caaagcacac ctaaataatg atgaacaaat
     7201 caagacctac aagaaatgaa cacatagatg ctttttaagt gtcattagtt atcggaatca
     7261 aagaatttga ggtggtgaca actgggg
//
