LOCUS       Exported                6914 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor MYNN with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pMYNN-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6914)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6914)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6914
                     /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             3510..3530
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3543..3587
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3627..3650
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3678..4397
                     /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(3723..3744)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3984..4003)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4435..5020
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4601..4618)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4828..4847
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5102..5905
                     /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(5165..5184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5775..5794
                     /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 gattgctccc ttgagtagag cagaatttct
     2701 cacctctgaa aatatatgaa agtgccatac atactgttct gaactcacag aataatatga
     2761 acaacttatt aaaaacattc agtgaatgaa aagagtattt cttatcactg tttaatattt
     2821 tgagggagtt aaagaagtat aacgacattt actctcaaag aatttacagt ctggttgagg
     2881 attcaaggca tacctgagta gaatgttaaa tggcaataag agagatagat gccacaaggc
     2941 agtatatgaa aattgacaaa tgtgtgatta agccgatata acagctcacc ctgtttcatt
     3001 cttaatcaag tccatttttt taaatggagt tggtgcttga acttaacctt gaggcccaga
     3061 ttgaatttac atagatagtg aggagggcat atgtacaaaa gagaaaccgc ttgagtaaag
     3121 gtatggtggt tggaatgtaa tttctgggaa tggtaagtac attagtctac ctcaggggtt
     3181 tagttttgtt ttaaagataa tgtagatttt tttttccttc cattctaggt gcagataaaa
     3241 ctctagactc cagtgcagag gatcatactt tgagtgaaca ggattccata caaaaaagtc
     3301 ctttatcaga aactatggat gtgaagcctt ctgatatgac tttaccatta gctcttccac
     3361 ttgggactga ggaccatcac atgcttctgc ctgtcacgga tactcagtct cctacatcag
     3421 atacattgtt gaggtcaact gtgaatgggt attcagaacc acagttgatt tttttacaac
     3481 aattatacgg gattccaact actgcaagcg agaatcttta ttttcagggc gccgccaaat
     3541 tcaaagaaac cgctgctgct aaattcgaac gccagcacat ggacagcgga ggtggaggtt
     3601 cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt ccaggggccc ctgtcgtcga
     3661 gcggtccctc gggttcgatg gtgagcaaag gcgaggagct gttcaccggg gtggtgccca
     3721 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3781 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3841 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3901 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3961 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     4021 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     4081 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     4141 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     4201 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     4261 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     4321 agcgcgatca catggtcctt aaggagttcg tgaccgccgc cgggatcact ctcggcatgg
     4381 acgagctgta caagtaatga tctagagtcg agttaattaa gaattccgcc cccccccctc
     4441 tccctccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     4501 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     4561 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcag
     4621 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     4681 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     4741 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     4801 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     4861 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     4921 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     4981 gttttccttt gaaaaacacg atgataatat ggccacaacc tgttacattg cacaagataa
     5041 aaatatatca tcacgaacag taaaactgtc tgcttacata aacagtaata caaggggtgt
     5101 tatgggatcg gccattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5161 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5221 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5281 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5341 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5401 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5461 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5521 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5581 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5641 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5701 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5761 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5821 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5881 tcgccttctt gacgagttct tctgagtctg atgactttgt aaggaatatg gaattgctaa
     5941 gatatcattg gtagcaaaca tctctggtaa ggtgcatata ttcatattaa attcccattc
     6001 atttgagttg tgaccattat ttttcattca ctgaagtaaa agcacctgat gctgcatcac
     6061 aactgcaatg tttgttttta tttttggctt tatgtctata ctccacagag agctttttaa
     6121 gtaatgaatt atgtagcact attttgggtg gatgagtttt attttctttt aaagctgcct
     6181 taattccatt tttattttgc attttagaat tgcccttcat ttactaaatc agccagttaa
     6241 tgtgattcag ttgtgacctg tgggatttta aaattttcct gccctcataa tttaagcaaa
     6301 atccacgata gtatcagccc aagagtgtgt tgctatttgt aagccccatg gatcaataca
     6361 taaactatgc ataaaaataa aatttcatat tttcgctcgt aaaaatttag gcgctgaata
     6421 agaattgtgc tatttctcta ttaatatatt gttagaaata acttttctaa tgaaacaaaa
     6481 atgtttctca aaatctagaa aagtgcatat aaatttttgt tcatttagaa ctgagtattc
     6541 tggagaaaac caatggtcag cacttttggt ttgagttatc tgagattgta ttttcaaaat
     6601 gaattttaag tagatatttg gtgtttttat cttgtcaaag acagtttcaa aatttgattc
     6661 agtttatgtt ttattccgtt ctaaaatatt ggagatgagg aaaatgtact tgagttttga
     6721 aatattttgg taagcctctc ttactcctgt tatgcacttt ttaaaataaa tactgaattc
     6781 cagaaagaat tttctagata taaacttaat gcatactaga aatcacttgg cttctaaatt
     6841 tggtacatta tattgcataa agagttaaga aattaacctc aattataagt aaacattttg
     6901 agttaccaga aagc
//
