LOCUS       Exported                7113 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ESR1 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pESR1-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7113)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7113)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7113
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(36..52)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(36..52)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(49..71)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    60..76
                     /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(84..114)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(267..284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(438..1026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(518..537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1197..2057)
                     /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     1820..1839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2058..2162)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2230..2248
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2286..2308)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2408..2427
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2621..2643
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2635..2652
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2636..2652
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3702..3722
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3735..3779
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3819..3842
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3870..4589
                     /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(3915..3936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4176..4195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4627..5212
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4793..4810)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5020..5039
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5294..6097
                     /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(5357..5376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5967..5986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
       61 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      841 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
      901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2161 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2221 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2281 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2341 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2401 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2461 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     2521 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     2581 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2641 acgacggcca gtgaattcga gctcggtacc cgggggctga gcctgcagag gcttgaggct
     2701 gctgccactg ggtagcggta ggcctttccg aggaggcggc atttgaagac cggaggaagg
     2761 ttcatcccag caagtaggaa cagcaagtgt aggtccccta agtcttgggg gagcttagtt
     2821 cctttaaggg cagcacaaaa atcagtgtgg ctccggagag cacattaggg gagagaggca
     2881 ggaagagctt ggagacatgg atggaagctg gaccagttgg gccttgttga acatggaaag
     2941 gcatttagat cgtattctga gttaaatggg aagtgacgtg agagatttaa caatggagcg
     3001 tcttgaactg ctttactcat ttaaaatacc cactcctgct tggctgaata tctcatgttg
     3061 tctttttaga agctttggcg atcctatttg aatgcattta ggtcctattg gaggggaata
     3121 ggatctcatt tgaggccacg gaggtccatg gaagtcacct gcatagcaaa taccctgaaa
     3181 gtggctgcag ggagagtgtg agggtgggac cgccctggta ggaggtggaa aatgaaaaac
     3241 acacggccat gagttccaga ttagggcttc tgaaagccct cagctttccc agctcccatc
     3301 ctaaagtggg tctttaaaca ggaagaaaga aagattgcta agtgtctttg gagttcctct
     3361 tccttcccct tctagggatt tcagcactcc tggggctcgg gttggctcta aagtagtcct
     3421 ttctgtgtct tcccacctac agtaacaaag gcatggagca tctgtacagc atgaagtgca
     3481 agaacgtggt gcccctctat gacctgctgc tggagatgct ggacgcccac cgcctacatg
     3541 cgcccactag ccgtggaggg gcatccgtgg aggagacgga ccaaagccac ttggccactg
     3601 cgggctctac ttcatcgcat tccttgcaaa agtattacat cacgggggag gcagagggtt
     3661 tccctgccac ggtcgatccc gggattccaa ctactgcaag cgagaatctt tattttcagg
     3721 gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg
     3781 gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc
     3841 ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg
     3901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3961 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4081 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4141 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4261 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4321 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4381 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4441 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4501 accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca
     4561 ctctcggcat ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg
     4621 cccccccccc tctccctccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     4681 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     4741 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     4801 aaaggaatgc agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     4861 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     4921 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     4981 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5041 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5101 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5161 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa cctgttacat
     5221 tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa
     5281 tacaaggggt gttatgggat cggccattga acaagatgga ttgcacgcag gttctccggc
     5341 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5401 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5461 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5521 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5581 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5641 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5701 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5761 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5821 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5881 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5941 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6001 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6061 catcgccttc tatcgccttc ttgacgagtt cttctgagtc gactgagagc tccctgggtc
     6121 ccacacggtt cagataatcc ctgctgcatt ttaccctcat catgcaccac tttagccaaa
     6181 ttctgtctcc tgcatacact ccggcatgca tccaacacca atggctttct agatgagtgg
     6241 ccattcattt gcttgctcag ttcttagtgg cacatcttct gtcttctgtt gggaacagcc
     6301 aaagggattc caaggctaaa tctttgtaac agctctcttt cccccttgct atgttactaa
     6361 gcgtgaggat tcccgtagct cttcacagct gaactcagtc tatgggttgg ggctcagata
     6421 actctgtgca tttaagctac ttgtagagac ccaggcctgg agagtagaca ttttgcctct
     6481 gataagcact ttttaaatgg ctctaagaat aagccacagc aaagaattta aagtggctcc
     6541 tttaattggt gacttggaga aagctaggtc aagggtttat tatagcaccc tcttgtattc
     6601 ctatggcaat gcatcctttt atgaaagtgg tacaccttaa agcttttata tgactgtagc
     6661 agagtatctg gtgattgtca attcattccc cctataggaa tacaaggggc acacagggaa
     6721 ggcagatccc ctagttggca agactatttt aacttgatac actgcagatt cagatgtgct
     6781 gaaagctctg cctctggctt tccggtcatg ggttccagtt aattcatgcc tcccatggac
     6841 ctatggagag cagcaagttg atcttagtta agtctcccta tatgagggat aagttcctga
     6901 tttttgtttt tatttttgtg ttacaaaaga aagccctccc tccctgaact tgcagtaagg
     6961 tcagcttcag gacctgttcc agtgggcact gtacttggat cttcccggcg tgtgtgtgcc
     7021 ttacacaggg gtgaactgtt cactgtggtg atgcatgatg agggtaaatg gtagttgaaa
     7081 ggagcagggg ccctggtgtt gcatttagcc ctg
//
