LOCUS       Exported                7130 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZBTB40 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pZBTB40-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7130)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7130
                     /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             3713..3733
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3746..3790
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3830..3853
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3881..4600
                     /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(3926..3947)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4187..4206)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4638..5223
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4804..4821)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5031..5050
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5305..6108
                     /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(5368..5387)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5978..5997
                     /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 cggagaagta gctgggcttg gcttcctcac
     2701 ttgttcctgc tctctgctct tcactgattc tcaagagcaa ggtggctgtg gcctctagtt
     2761 taaaaatatc aagggcatct agctcttctg ggatctatgg gttagaaaca tcaactggga
     2821 ggcttaggcg ggaattggat atgcctaacc acagactcat tctctctcag ttggaaaggt
     2881 ggtctctaac agattatgca gatttggaag ggacataggt gggctgtgga ggagagaagg
     2941 gggcactggc cccgccaggc tggtgagctg aatgaatgcg tggagtcaca gttcatccag
     3001 tcttcctcct aagctcggtc actgtgtggc ctcctgggaa ggctgtgagc ctgcgagagg
     3061 tccagagagg ctgccccggg cccacctggg cctgtggtgg atcaatccca ttcccatgtt
     3121 ccaggcatga gaccagctgg cttcagtggc attcctctca catggcactc acctctgcag
     3181 tgtcatccca agacaggaaa gcccagtgtg aaaaacagag ccagtgttcg ctggtgtata
     3241 atcatagagt ccgggcgccc cgagatggcg tactgagcaa aattatagca atttagtgca
     3301 cagttattta aattgtttgc tggccttctc ttttgcatcg ccagtgggtt agacttgcct
     3361 cttcccatta ggcacatgta agtgctctcc tcaggtgaaa acataaatat catcattaca
     3421 gcatgagatg aaaaccatgt aaatcaggga gccaactgtg aacactgccc agagcagcct
     3481 cacggtcttt ctctttcagg tgatccaaac cccagagccg gtggccccga cagagcaggt
     3541 gatcactttg gaggagaccc agcttgccgg gtcgcaggtg tttgtgacgt tgccagattc
     3601 tcaggcatct caggccagct ctgagctcgt ggcggtgact gtggaggact tgctggatgg
     3661 cacagtgacg ctgatctgtg gtgaggccaa agggattcca actactgcaa gcgagaatct
     3721 ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca
     3781 catggacagc ggaggtggag gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct
     3841 gttccagggg cccctgtcgt cgagcggtcc ctcgggttcg atggtgagca aaggcgagga
     3901 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3961 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     4021 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     4081 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4141 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4201 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4261 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4321 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     4381 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4441 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     4501 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc
     4561 cgccgggatc actctcggca tggacgagct gtacaagtaa tgatctagag tcgagttaat
     4621 taagaattcc gccccccccc ctctccctcc ccccccctaa cgttactggc cgaagccgct
     4681 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4741 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4801 cccctctcgc caaaggaatg cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     4861 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     4921 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     4981 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     5041 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     5101 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     5161 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     5221 acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact gtctgcttac
     5281 ataaacagta atacaagggg tgttatggga tcggccattg aacaagatgg attgcacgca
     5341 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5401 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5461 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5521 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5581 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5641 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5701 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5761 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5821 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5881 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5941 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6001 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6061 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt ctgagcagcc
     6121 tttcatccgg cagagccttc ctgcgtttgc agcagagagg aggccccaca gcttgccctt
     6181 tgccctccat ccctggctgt cctgagtggt gagcatctta gcttagcacc aaccaacaca
     6241 gtctcaccta gaaaacagat ggaagcttcg ttgttctcat agaaccaaca gcatctgagc
     6301 cctcaacacc aacagcacca tcctctgtag cagacaggcc tccctcccca caggcccgct
     6361 gctgcggcct ctatgacact gtccatcccc aagtgacatg tggcttcaga agtagagtct
     6421 gaaagagcag tgggatggga gctggtgacc agggtacccc aggaggcatg atgtgccgac
     6481 agcgctcagt gggcagaatg gcagttagat atgggacttg gcccacagtg ggggctgcaa
     6541 atgctgccac tgcctctggc catttaaagt gagaggggca ccaacagaca attcggggac
     6601 cttaggcccc ttcctgaggc acacttggcc ccttcctcgg tcctatcatc ctaccctctg
     6661 ctggggttcc ccctccaccc agtggccacg cccagcaccc tattgatggc tataggacag
     6721 gtagccctca tttccatgcc tgataacccc ttgtcagttg tcagctcttc ccaaaacaaa
     6781 gctctggagt ttctgctgga agtgtttaat gtgaggcatc agctgctaga cagtgtctgc
     6841 ctttccagga cattcttctt ggtattcctt acctgaagcc tggttcccac agctcttccg
     6901 tgtcatcttc atccccttcc tttaatcaaa ggctgaaatc tctgcctgac ctggcagcct
     6961 ggctccctct gggtcatatg gtgcagactg tgaccagcac caggcaggca gtcctgtctg
     7021 cgtgtggagg agcagccgtg actgcccgtg gctctgctgc cgcccactcc ctgccttgtg
     7081 agtggcctgc tgcctcacct cccgcaggcc gccacactta ttgcaggtca
//
