LOCUS       Exported                6944 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor DLX1 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pDLX1-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 6944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6944
                     /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             3590..3610
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3623..3667
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3707..3730
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3758..4477
                     /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(3803..3824)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4064..4083)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4515..5100
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4681..4698)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4908..4927
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5182..5985
                     /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(5245..5264)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5855..5874
                     /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 tagaagaaca 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 acctaggtcc 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 tcggtacccc ggctttctgt aagactccgg ggagcccgtg
     2701 agcgttcctg acggcggcgg gcgcgggttt ccgacgtccg gtccgggatt tggagcagag
     2761 ctggagaaag caaacagaca ctagtgcaag aatggttttg aatccaaaga gaagttcagc
     2821 aaaaccttga ggcctcctta gtccgtccca actcaagggc agaaaagatg gcgcatagaa
     2881 aagttgggtc gcgttgcaaa taaatttcct ccactccttc ctaggttttc gattcttttt
     2941 attgatgttg atattggaat tgctggctcg gtgttatgca ggcgctgtat cttccgcagg
     3001 cggtagccac ggtcggactg agcccctggc aggctaaggc ctggatcccg gttctcccct
     3061 catcacaact cctccctggc tctctgagag ctgcccccgt gggctgacgg cggcgggcgg
     3121 ttcggccttc gagcctttgg ctcttatggg ggaagggagg aaggagggat gtctctgctt
     3181 ctctggcagg gagctgccag ctggcgcagg gttgggaggt cctatctctg ctgttctgcg
     3241 gtcccttttt tcctccctgt gacctaggta ggctcagtgg tcccagcctg agtcacgttg
     3301 ttgtccgctc ttgcaggtca agatctggtt ccaaaacaag cgatccaagt tcaagaagct
     3361 gatgaagcag ggtggggcgg ctctggaggg tagtgcgttg gccaacggtc gggccctgtc
     3421 tgctggctcc ccacccgtgc cgcccggctg gaaccctaac tcttcatccg ggaagggctc
     3481 aggaggaaac gcgggctcct atatccccag ctacacatcg tggtaccctt cagcgcacca
     3541 agaagctatg cagcaacccc aacttatggg gattccaact actgcaagcg agaatcttta
     3601 ttttcagggc gccgccaaat tcaaagaaac cgctgctgct aaattcgaac gccagcacat
     3661 ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt
     3721 ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag gcgaggagct
     3781 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3841 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3901 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3961 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4021 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     4081 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4141 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4201 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     4261 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4321 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     4381 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc
     4441 cgggatcact ctcggcatgg acgagctgta caagtaatga tctagagtcg agttaattaa
     4501 gaattccgcc cccccccctc tccctccccc cccctaacgt tactggccga agccgcttgg
     4561 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     4621 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     4681 ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     4741 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     4801 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     4861 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     4921 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     4981 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     5041 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     5101 tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata
     5161 aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt
     5221 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5281 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5341 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5401 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5461 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5521 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5581 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5641 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5701 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5761 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5821 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5881 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5941 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtctg aggttgcccg
     6001 cccgtctcct tcttgtctcc ccggcccagg tccctcccgc ctccaggtcc atccatcccg
     6061 tccggaaaag aaggacccag agggaagaag gaacagtgga ggcgggacgc cctccatctc
     6121 ctcggagccc cgcgaggtcc ggcccagcaa cttcccggca tccgcgctct agcctgaacc
     6181 ctggcctggg ccgagcagtg gcagcagaga gtggcctcgg agggaagcca ctgccacctg
     6241 agacagccca agcagcaaga taaacccgct ccacccgacc cgccgacctt cagctttgtg
     6301 ggactatcag gaaaaaacaa aacaaaaaca aaatgtagaa aaagcaaaag ctcttttctg
     6361 tcctgtcagt ctcctgtctc cttttgctct gtctgtgcgc tggtaaagtc caggtcctca
     6421 tccgtccgct gtcctcattc tgcagcctca gcaaaaagcc acaagatctg agcggcccgg
     6481 gtcctgccgg gctgaccatc tccggatcct gggacactct gcctgaccat ctgtgtagct
     6541 ggtgtgggaa tctgggggca ttggagggag ggggttttat ttattgagaa atggacttcg
     6601 cctgaggctg tttgccaatt cagggttctg ctgggcgcaa ggaacgcact gttcaaacgc
     6661 actgtttact ttaagcgcac ggggagaaac gaataaggag gacgtggtga tttttaattt
     6721 atacagtaac ttttgtactt ctctggtatg gagagtttgg agccgaatga tttgcatttt
     6781 ttacatgtcc gacattattt aataaataat ttttaaaaga aaagaacgat aaatgaagcc
     6841 aacatgattt tctcatttcg ggaggaactc tgttgcttcg cctggacaag aaggaaaatg
     6901 ctgatttcct ccttgggtag aaagagggag cgagggcaaa tggg
//
