LOCUS       Exported                7387 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor NR2F1 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pNR2F1-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7387)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7387)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7387
                     /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             3876..3896
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3909..3953
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3993..4016
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4044..4763
                     /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(4089..4110)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4350..4369)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4801..5386
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4967..4984)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5194..5213
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5468..6271
                     /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(5531..5550)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6141..6160
                     /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 tcggtaccca actccccacc acccacatcc ccccctaccc
     2701 ccccagttct gaagcagcag cagctgctgc ctgtagccta gctttgtgtc tgattgctga
     2761 agatgtcgcc tttctgcagc ctggagtaac tgagtcatat tcttcctgat tattatacag
     2821 cttacagaga tgtgaccaat tttcaattac ttgatctaag gtatagatag aagtgtacac
     2881 cctcaccaag gatttgctgc agagtccttt cttgcataga taacggatct gaaatcctat
     2941 attgctaaag cgtgggcttc atctgattcc aggcactgcc agcccatgat tccttacccc
     3001 ctcccatgtg tttgttcact acacaatcta ggttctcctg gaggtttctg gttggggtgg
     3061 tttgggtttg agaggggact ccaggaatcc tttaagaact gccttcaggc tgcatatttc
     3121 ctaaggggta ctgaacaccc ccagatcaga ggcaaatggg gcaaaagtta atgagcacgg
     3181 ccagaaagat gctcccttgc aggccgagga caggttgtta agcgcagcaa acatggttcc
     3241 agcattgtac attatttaat ccaaaggtga aagaggaaaa acaaagggac tagagaggtt
     3301 tctgcctctg catgtgtgtg cctctctctc cagctcccct aggcttggtg ggggtttgat
     3361 ttatttttat ttgtattgta ttgggggcgg gggaggaggg aatgaagaag gggatggaga
     3421 ggtataggag ggtgaatttt tcttttctct tttacttctt ttttattttc cattttctcc
     3481 ttaaaaaaaa ttgatggctt attttgcctt tgctatttgt cagcctaacc gtgtgctccc
     3541 tttccctgtc tctccctcct gtggctgctt gggcagacgc ctgtggcctg tcggatgcgg
     3601 cccacatcga gagcctgcag gagaagtcgc agtgcgcact ggaggagtac gtgaggagcc
     3661 agtaccccaa ccagcccagc cgttttggca aactgctgct gcgactgccc tcgctgcgca
     3721 ccgtgtcctc ctccgtcatc gagcagctct tcttcgtccg tttggtaggt aaaaccccca
     3781 tcgaaactct catccgcgat atgttactgt ctgggagcag cttcaactgg ccttacatgt
     3841 ccatccagtg ctccgggatt ccaactactg caagcgagaa tctttatttt cagggcgccg
     3901 ccaaattcaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac agcggaggtg
     3961 gaggttcgag cggtccctcg ggttcgtcga gcctggaagt tctgttccag gggcccctgt
     4021 cgtcgagcgg tccctcgggt tcgatggtga gcaaaggcga ggagctgttc accggggtgg
     4081 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     4141 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     4201 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     4261 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     4321 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     4381 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     4441 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     4501 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     4561 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4621 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     4681 acgagaagcg cgatcacatg gtccttaagg agttcgtgac cgccgccggg atcactctcg
     4741 gcatggacga gctgtacaag taatgatcta gagtcgagtt aattaagaat tccgcccccc
     4801 cccctctccc tccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     4861 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     4921 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     4981 atgcagggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     5041 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     5101 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     5161 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     5221 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     5281 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     5341 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaacctgtt acattgcaca
     5401 agataaaaat atatcatcac gaacagtaaa actgtctgct tacataaaca gtaatacaag
     5461 gggtgttatg ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5521 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5581 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5641 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5701 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5761 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5821 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5881 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5941 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6001 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6061 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6121 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6181 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6241 cttctatcgc cttcttgacg agttcttctg agtcaccttg ggcgcttccc acctgccccg
     6301 tccccctaga gactcagagg acccacctgg gccaaggact ccaaagccgc ggggacaccg
     6361 ggaagtgcag cgggccaggc aggctgggtg ggagggagga gggccgagac aggagcagcc
     6421 cacccagcag aaatacaatc cgagctacaa agcatgggaa aaagagactc ttttaggatc
     6481 agatctgtga gcacgttggc gaggaaaaac aaaacaaaca aaaaaaagaa ccttgtgtct
     6541 gtctggtgaa aaaaagaaaa acaaattgga agagaggacc atgagaattt taataaaaca
     6601 gaaggaaact aatggacctt ccaggattta ttgtggacgg atgtggatat attctgtaca
     6661 ggaacaacac atatggaagt ggactgaagc ctatgtagaa acacacacac actgaacatt
     6721 gttattcatt ttgtaaaata ctagtcttta ttttcatttt ttgtaaaatt taaacatcgt
     6781 atgcgcataa agaaaaagga aacaagaatt aggggaaaat aacattttcc aaataattat
     6841 aaaaaattgt cctgtgtcta tgtatctata tctgttttgt atttttttct ggttccaaac
     6901 cagatttcct gtgattctat actaataatt tttgatataa ccctttgctt cttataatga
     6961 gtgcgatata tgttgtcgag gctgttcttc aagaattaaa attgaagtga aaatttaaac
     7021 aaaaataaaa gaatttagca aaacccatgt gtctggttgc ttgatagatg ttatccgtgg
     7081 aagagaactc cttggttaat ttccacctca actattatta gaattgaagg ttttggtgtg
     7141 taaaagggga gacttttaaa gtctactaca aagttggtta gattcatttc caaaaaccaa
     7201 gctgctacct tggttttgta aatgcggagg gccaggctct acatgggctg gggtccgcgg
     7261 ggaggcaggg cgccttggtc gctgacttca gccgttgaga cctggtgggc tgcgggcggt
     7321 cccgtgctca gttgggccag ggagtggtgt aggcagggtc gcgcgcccgg taagtctcca
     7381 ggcacca
//
