LOCUS       Exported                7286 bp ds-DNA     circular SYN 12-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 7286)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7286)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7286
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1250..1270
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1283..1327
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1367..1390
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1418..2137
                     /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(1463..1484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1724..1743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2175..2760
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2341..2358)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2568..2587
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2842..3645
                     /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(2905..2924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3515..3534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4687..4703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4687..4703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4700..4722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4711..4727
                     /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(4735..4765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4780..4801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4918..4935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5089..5677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5169..5188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5848..6708)
                     /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     6471..6490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6709..6813)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6881..6899
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6937..6959)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7059..7078
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7272..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7286..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtaccctggg agaagggagt ttgagaaggg
       61 aaagtgggga aggaaaatta aagaggtttt aggaaagagc acatatggga gttgaagaca
      121 cggaagtcag ttctcctgta tgcacatatt tgtgtccgtg tgctccttgg agagtcttgg
      181 ggttaaccca gggaaatgca taccaggctt taaaacaact cttccaatct gcttagtttt
      241 ttaatattta aaaaccaaat gggaggggag gaaaaaactt ttatgatcct agatattcaa
      301 gaaatacatt atttgtgacc tgtaaagcct cccatataaa tccctctgga tgtggggtag
      361 tccttaggga cacaggaata taaaggggtt gtttttcatt taaataatca caaattttct
      421 tgcatttttc agatgataca gaactttcag gaagaagcat gttttattct ggacacaatg
      481 aaaggtaata gtaagacaaa catctttctc taagcagttt aacagcttta ctactactag
      541 gcaaagtaca tacttctttg aggagagata ggaaaagtaa ccatatttgg gtaaagaaag
      601 tgacaatatt agagattgtt ccttttttta agttggcttt tttttttttt ttttttttga
      661 aacagagtct cgatctgttg cccaggctgg agtgcagtgg catgatctcg gctcactaca
      721 accttcgctt cccaagttca agtgattctc ctccctctgc ctcccaagta gctgggatta
      781 caggcacgtg ccaccatgtc tggctaattt ttgtattttt agtagagatg ggtttcacct
      841 tgttggccag gctggtctcg aactcctgac ctcaagcgat ccacccacct tggcctccca
      901 aagtgctggg attacagaca tgagccactg tgcctggccc agctttttgc cattctcttc
      961 catcttccat tacagatgag gaaattgagg catgttgttt ttataatgca ttttttttga
     1021 gacagaattt cgctcttgtt gcccaagctg gagtccagtg gcgtgatctc agctcactgc
     1081 aacctccacc tcccttttct ccccttctca cgagaatctt accatacttt ttttctcctg
     1141 ctgtgtttta gaggtgacta aagatgattc tctctctctc ttttttttaa cagcagaaac
     1201 caatgatagc tatcatatca tactgaaggg gattccaact actgcaagcg agaatcttta
     1261 ttttcagggc gccgccaaat tcaaagaaac cgctgctgct aaattcgaac gccagcacat
     1321 ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt
     1381 ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag gcgaggagct
     1441 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1501 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1561 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1621 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1681 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1741 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1801 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1861 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1921 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1981 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     2041 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc
     2101 cgggatcact ctcggcatgg acgagctgta caagtaatga tctagagtcg agttaattaa
     2161 gaattccgcc cccccccctc tccctccccc cccctaacgt tactggccga agccgcttgg
     2221 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2281 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2341 ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2401 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2461 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2521 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2581 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2641 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     2701 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     2761 tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata
     2821 aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt
     2881 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2941 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3001 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3061 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3121 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3181 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3241 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3301 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3361 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3421 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3481 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3541 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3601 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtcga gtgcggcgtt
     3661 tcgtgcccag tggctgctcc ttccttcacc tctgaaaacg gccctcttga agggggatat
     3721 gaatggagat ttgaaggtct gcaagaacct gactcgtctg actgtgtgtg gaggagtcca
     3781 ggccatggag gcagaatcct ggccctctgt gttggcccaa gctcttgtgg tacacacaga
     3841 ttactgccca atatgcagtt ctgcagctgt tttagttaaa tttctggacc ttgttgttgt
     3901 taaatatcag tagaaactct acataattta gagtgtatgt agggcataat gatgatggga
     3961 attgtgtgat gtttaacagg aagatcttaa attttgtgat atggagccct gtaatttttt
     4021 tcttatataa aaatgggtat ctatattcat aagactaggt cttcaatctc tttgtactgg
     4081 tctcaaatgt actggtatct ctgctttttg ccacagtttg tccctgaaaa ctttatcagt
     4141 gaggagaaat acagaatttt ccttttggtt cccctgtaat accacaacta atagtatttt
     4201 cagctaacat ttattgacca ggcagtgtga taaatttttt gaatgaccat cttgtttaac
     4261 cccctaatca cactatctga agtaggtgct atagtgaccc ccaaccgaag atgaggaaat
     4321 ggagacacac agtggctaag tagcttgcta ggtgccagga gctggcaggc agtgatgctg
     4381 aaatccaaac caggcaatct ggccctcctt attcaccctc tcataccacc agtgccaccc
     4441 attataacct gtgttcactt cgcttattac tgtggccctt ctggtgacta caacataaca
     4501 ctgagtatga actgaggaat taaccactga aagagacaaa gcacacctaa ataatgatga
     4561 acaaatcaag acctacaaga aatgaacaca tagatgcttt ttaagtgtca ttagttatcg
     4621 gaatcaaaga atttgaggtg gtgacaactg ggggacctgc aggcatgcaa gcttggcgta
     4681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5701 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5761 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5821 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5881 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5941 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6001 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6061 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6841 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6901 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6961 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     7021 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     7081 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     7141 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7201 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     7261 gccagggttt tcccagtcac gacgtt
//
