LOCUS       sequence-214773-        7419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor GLIS3 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7419)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGLIS3-donor
FEATURES             Location/Qualifiers
     source          1..7419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(82..88,89..106,107..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(join(1195..1986,1987..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             3875..3895
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3908..3952
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3992..4015
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4043..4045,4046..4048,4049..4762)
                     /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(4088..4109)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4349..4368)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4800..5373,5374..5376,5377..5385)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4966..4983)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5193..5212
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5467..6270
                     /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(5530..5549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6140..6159
                     /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 cactaccccg attcccagag aattgtgccg
     2701 cctgtcagct cctgcagtgt ggtgccttcg tttgaggact gcctagtccc tacatccatg
     2761 ggccaggcca gttttgatgt tttccacaga gccttctcga ctcactcggg cattacaggt
     2821 gtgttggttg tctgtgactt agaaaatcct gtcaactgac aggagagacc aggtgggctg
     2881 cccgtgaccg acctgggctg gacgtgacca agcacatgac cgcatcaaat cctgttgaac
     2941 gacccctgaa atggctctca tctccacctt cccctccagc cctccttcag cccctcttct
     3001 tccaccaccc tggtgcagga gccatcatgt ctctcccaga ctatggccat aaccttccag
     3061 taagtctccc tgacctgatt cctcatcttt accattctgc cctccatact accatgaaca
     3121 ttgcctcccc aaaatgaagg ataaaatatg cctcttttct gctcaaatat attaaatagt
     3181 ttccaattgc ctagagaata aattcgactc tccagcatgg cattcaaagc ccccataagt
     3241 taggcctgat ccagctctgc acacctgccc acacatattt ttccctacat gctcttcacc
     3301 tgagctttgc cagcaccctt gaccccacat gcctctccct ctctgtcttt gctctttgtt
     3361 tcctccttct ggaatgttct ttccctattt cttcatctgt gagaattcca ttctctttca
     3421 tgactcaacc ttaacaccag ctcctttttg aggccacccc cagccttccc tagaacactt
     3481 ttcttttact tcttctcagc ctgtcttttg tgtggtcata ttttagccag ctgcctgcaa
     3541 gtcagcctct tccctcacta aggtcgctgg aaacagagac tcactatagg gcccaggctg
     3601 gcatggttgc ctccttagcc cccagtcctc acttggtgag ctgcattact gtagtgcatt
     3661 tccagtgggc gtggcatggg gcaggagtta ctgacttaac tgcgttcttt ctcttccagt
     3721 gtatgattta ccttcaagtt cctcgagcct ctttggggag tctctccgca gcggggctga
     3781 agatgctacc ttcttgcaga tcagcaccgt ggaccgctgt cctagccagc tctcctctgt
     3841 ctacaccgaa ggcgggattc caactactgc aagcgagaat ctttattttc agggcgccgc
     3901 caaattcaaa gaaaccgctg ctgctaaatt cgaacgccag cacatggaca gcggaggtgg
     3961 aggttcgagc ggtccctcgg gttcgtcgag cctggaagtt ctgttccagg ggcccctgtc
     4021 gtcgagcggt ccctcgggtt cgatggtgag caaaggcgag gagctgttca ccggggtggt
     4081 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     4141 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4201 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4261 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4321 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4381 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4441 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     4501 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4561 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4621 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4681 cgagaagcgc gatcacatgg tccttaagga gttcgtgacc gccgccggga tcactctcgg
     4741 catggacgag ctgtacaagt aatgatctag agtcgagtta attaagaatt ccgccccccc
     4801 ccctctccct ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     4861 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     4921 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     4981 tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     5041 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     5101 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     5161 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     5221 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     5281 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     5341 acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta cattgcacaa
     5401 gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag taatacaagg
     5461 ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5521 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5581 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5641 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5701 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5761 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5821 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5881 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5941 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6001 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6061 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6121 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6181 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6241 ttctatcgcc ttcttgacga gttcttctga gtcaagctct cttggccact cctgcacacc
     6301 caggaccttg atgttgcttg ccaccttttg ttttcgtact ctcacagact gcttgaggaa
     6361 ggatgtcagc ccaatcagcg ggccctgcag agttactgat gttctttcag aaggcaggac
     6421 tgaatggcag agctagcttt gcaggaatca ttacagttgc tcctttttct ctctggtttt
     6481 cctggactat gtgaccagga ggactgtctt ttcaaaggga atttagagcc tcactctcct
     6541 ggatacctgt tacttcctgg ctgtcaaccc ttaaaggacc ccagtgaaat gatgtgcata
     6601 tgaaaatgta gtttggggat gactctacct gaaaactcca aacagggaaa aagacctcaa
     6661 ttacgagagg catagtgacc tgcctcacct aatcctcacc cttgctgctc ctgatgcaaa
     6721 gccaggcaca acaagggaaa tccatgtcca aagcatagtg tggtctcaac tcttagaaac
     6781 tccctgggtt tcccttttct cttagccaca tgctaaccag ttgcttctca gtgtttctct
     6841 tcaatagaaa aataaggttt tatatgccag cagcctatcc ttgtttttgc attttaaaga
     6901 cttagttatt tatacgtatt ttaaaatgaa aaaaaaatgt aaaggtctgt gctttggatg
     6961 gaaatcatta ttattgtaat caccccagct atagacattt tgcacagctg tggttctgcc
     7021 tggccccacc tctagcatgt tagttttgca gttagcaaag gttacctgag ggaaatggag
     7081 tgggcacctc ccgcaccaac agtcctgtag ctggaacctt cctgagttga ctgtttagta
     7141 actccacctc aaagactgtg ttcaaagaaa aatgttgttt gactggttta cccattgcct
     7201 ccactctggt gctgcaccca gagaagtgcc agatgtgaga actgggacag actgagtcat
     7261 gtgccagctt cccttggaaa accaaaccta gtgatacttg acttccagtg ctcagatttt
     7321 actaacgttt ttcaggatat ccactcatct gtctgtctct tccctacact tctctccacc
     7381 ctctttttgc atccctctta ccctcttcct gactgtggt
//
