LOCUS       Exported                7104 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor SRF with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pSRF-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7104
                     /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             3656..3676
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3689..3733
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3773..3796
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3824..4543
                     /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(3869..3890)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4130..4149)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4581..5166
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4747..4764)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4974..4993
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5248..6051
                     /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(5311..5330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5921..5940
                     /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 attcctaggg acggaatggg agttatcagt
     2701 ggggaccagt gccgagctga cacatgtctg tgtttgccag tgacgctgcc cgccaccatc
     2761 atgacgtcat ccgtgcccac aactgtgggt ggccacatga tgtaccctag cccgcatgcg
     2821 gtgatgtatg cccccacctc gggcctgggt gatggcagcc tcaccgtgct gaatgccttc
     2881 tcccaggcac catccaccat gcaggtgtca cacagccagg tccaggagcc aggtgagtag
     2941 aggagcaggg ctaaggaaag gaggaccgtt tccttcttta tacacacaca cacacacaca
     3001 tacacacaca tatgcactga tgcctacaaa tatttctacc caaatacaac acagacttgg
     3061 atgctcacac acacatttgg acaccccact tggacactca cacccgcatg caccctcaga
     3121 cacactggca ccctttccct tgggctctta catctgagac tgccccagtc acttgtgcat
     3181 ttgacaccac tcctccaata tctggaagtt tcaacaaaca atttgagtat ctcctgtggt
     3241 tttccaatgt aggtggcgtc ccccaggtgt tcctgacagc atcatctggg acagtgcaga
     3301 tccctgtttc agcagttcag ctccaccagg taggtaggga tatctttcac cccatcccag
     3361 atagccactt ctttgtcttg accttagggg atcctgttac cagttcatca aagccaaaat
     3421 ccctagcctg gaagcccatc tgcttttctg ctcaggcctg tggttggcag gcagggaagc
     3481 caggggagcc tgaactggct ggccagtccc tgcccctcct catacatccc cttccctcca
     3541 gatggctgtg atagggcagc aggccgggag cagcagcaac ctcaccgagc tacaggtggt
     3601 gaacctggac accgcccaca gcaccaagag tgaagggatt ccaactactg caagcgagaa
     3661 tctttatttt cagggcgccg ccaaattcaa agaaaccgct gctgctaaat tcgaacgcca
     3721 gcacatggac agcggaggtg gaggttcgag cggtccctcg ggttcgtcga gcctggaagt
     3781 tctgttccag gggcccctgt cgtcgagcgg tccctcgggt tcgatggtga gcaaaggcga
     3841 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3901 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3961 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4021 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4081 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4141 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4201 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     4261 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     4321 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4381 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4441 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtccttaagg agttcgtgac
     4501 cgccgccggg atcactctcg gcatggacga gctgtacaag taatgatcta gagtcgagtt
     4561 aattaagaat tccgcccccc cccctctccc tccccccccc taacgttact ggccgaagcc
     4621 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     4681 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     4741 tttcccctct cgccaaagga atgcagggtc tgttgaatgt cgtgaaggaa gcagttcctc
     4801 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     4861 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     4921 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     4981 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     5041 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     5101 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     5161 acaacctgtt acattgcaca agataaaaat atatcatcac gaacagtaaa actgtctgct
     5221 tacataaaca gtaatacaag gggtgttatg ggatcggcca ttgaacaaga tggattgcac
     5281 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5341 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5401 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5461 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5521 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5581 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5641 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5701 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5761 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5821 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5881 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5941 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6001 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agtctgatcc
     6061 gcccgccgcc ctggacagat ggcccaaggg atggcaccac ttatttattg ttgccttttc
     6121 acgttttctt tacacacacg ttgacgggcc gcaggaggga ggcggggagg aggaacgggc
     6181 agccacagga ctgagccctc tcactccagc caaagaaatg ggcctgcctg cctccacccg
     6241 tcctccctca gcctcccctt cttcccgccc cacctcccat ttctgttgct ggaggggctg
     6301 tcctccttcc tgggaccccc tcgccagctt ggctcgatgt ttgccatgag tattagctta
     6361 cccaatggga ccgtgcccca cctccccaca cacaggcctt ctgtggggct gggcaccgtg
     6421 tcctcctctg aggaagcagt tggggccctc ttgccagcct ccttgctgac cccaggtcag
     6481 ccctgtgtct gtcacaggct gggtcaaaag agccctggct ctgcccctca gggggccagc
     6541 tggggagatg ggggcttctt cctcacactg ctgtcctctc ccccttcagc tcctgagtag
     6601 ctgggcctgt gcactgggca ggttcctggg gccgcctgcc ctgccttgcc gctccccttg
     6661 gacctccagg ggctcctggg ttggagggaa ccaccagcgt tcccttctcc cccttgtctt
     6721 cccccctctc ctcccagctg ctttacttaa agttgatttt gaacttttta tttgaggaga
     6781 cgaagtgaaa acaaatctat aaatatatat ttttaaaata tttaactttt ttttatggcg
     6841 tttttctcgt ccccctccct gcccaaactc cccttccctg gggagccctc aggctcccca
     6901 gaactggctg ggcccctggg gacagagcca ccccatgagc tcggggtcca ccagtgtgtg
     6961 ggggagattc tgggtttgcc cagtcctggg ttgtttccag gagaaagccg ggggaggggc
     7021 cctcaggcca ttccccaacg gggtggggag ggtgacccac agctctgggc ctctttttgc
     7081 cctttagggc tgttgctagg gaga
//
