LOCUS       sequence-211724-        5066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor THRB with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5066)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 5066)
  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: pTHRB-donor
FEATURES             Location/Qualifiers
     source          1..5066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      31..619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     520..539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1674..1694
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1707..1751
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1791..1814
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1842..1844,1845..1847,1848..2561)
                     /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(1887..1908)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2148..2167)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(2599..3172,3173..3175,3176..3184)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2765..2782)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2992..3011
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3266..4069
                     /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(3329..3348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3939..3958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
       61 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
      181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggggaa gcccagtagt
      661 ctgcacagga cttgggaaga tttgctgggc tttccagggt cccggtgtct tctcactcca
      721 gctcccacta tagctgttct cttctctgca tggggtgact ctggcacagc tcccctcccc
      781 ttttcatttc aattccaaaa tcatgactaa tacaaggctg ctactggtag caattttact
      841 aaaggcataa gggaagggaa aggccagttc attgcagcct ggaagacaac acaaaattaa
      901 acatggtgtt tcttggtcct caaggaaatg tcttactatt ccgcgttcct cctttggcct
      961 caggcctgct gcaatgtccc cagtcccttc atatatctcc cccttctatg catgtccacg
     1021 ctcatcctct cctctctgcc aggcagccag tgaaaattca aaattgttcc cctcatcttg
     1081 cagaatattt cagctatgtc aggcttctcc ttctgtaccc taggagggag gcatcacctt
     1141 tcacactcaa gaaggtaatt cacactcaag ctaaactgtc taaacagaga tatttatctg
     1201 aacaaagtac tgcctctctt catataaatg atgccactcc cctggtacct ggtccctgga
     1261 gcaccagagt tcacgctgca tccatgctca tggctaagag ggaagaccct agatctgccc
     1321 gccccccaat aaaggcctgg aattggacaa agcaagcctt cccccttcca tctctgaatc
     1381 aatgtccatc ttctctcttc cccgcagatc gcccggggct tgcctgtgtt gagagaatag
     1441 aaaagtacca agatagtttc ctgctggcct ttgaacacta tatcaattac cgaaaacacc
     1501 acgtgacaca cttttggcca aaactcctga tgaaggtgac agatctgcgg atgataggag
     1561 cctgccatgc cagccgcttc ctgcacatga aggtggaatg ccccacagaa ctcttccccc
     1621 ctttgttctt ggaagtgttc gaggatgatt atgatattcc aactactgca agcgagaatc
     1681 tttattttca gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc
     1741 acatggacag cggaggtgga ggttcgagcg gtccctcggg ttcgtcgagc ctggaagttc
     1801 tgttccaggg gcccctgtcg tcgagcggtc cctcgggttc gatggtgagc aaaggcgagg
     1861 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1921 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1981 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2041 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2101 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2161 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2221 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2281 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2341 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2401 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2461 ccctgagcaa agaccccaac gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg
     2521 ccgccgggat cactctcggc atggacgagc tgtacaagta atgatctaga gtcgagttaa
     2581 ttaagaattc cgcccccccc cctctccctc ccccccccta acgttactgg ccgaagccgc
     2641 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     2701 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     2761 tcccctctcg ccaaaggaat gcagggtctg ttgaatgtcg tgaaggaagc agttcctctg
     2821 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     2881 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     2941 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     3001 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     3061 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     3121 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     3181 aacctgttac attgcacaag ataaaaatat atcatcacga acagtaaaac tgtctgctta
     3241 cataaacagt aatacaaggg gtgttatggg atcggccatt gaacaagatg gattgcacgc
     3301 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3361 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3421 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     3481 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3541 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3601 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3661 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3721 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3781 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3841 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3901 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3961 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4021 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ctgactggat
     4081 tcattctcat aattcctaca gcactactgg gtgtcatttc attccattgc ctagctcttt
     4141 tttgtttgtt tctttgtgtt gggagggatt atttgggagg gaaaagggaa gtagtccttg
     4201 gcatagacat ggatgaaatt gccccttgaa tgcgggtact tgaaactatt gcatttcgtt
     4261 ctccggtcct gtgatgtgaa tgctctgaag gttttatggt tgtggaggtg gggtggggga
     4321 caatcattaa ctcaccagca ccaagcatca ccagctccca cccgtccctg gtccaagact
     4381 tgagtcagca aaatggcgcc acaggacact aaagaagcct taaaaccaag ataatacgac
     4441 cacctccacc caatcctgat gttcgcaggg ctgaagttaa cagagcacag accaccttta
     4501 gttagatgtg gctttcagcc ttttagggaa agactcgaac aaattttcat ctattcaaga
     4561 gcatgctgtt tgcagcactc tttatggtct cctaggagca taacttggaa tgtacaggaa
     4621 tatttgtgtc ctaaaaaaat cagctgccct tccaatatca tttttataac gttatggttt
     4681 aattcaagtt atacaaaggt gtgactcaag gatgtgcagg tattgactga tggaggccaa
     4741 caaggaaaat attttttttc agtgagccca gagaaattag aaatcttcac caatgtcagt
     4801 gactgcatgg tttccaaaaa ttcacatagt attacctcac agtaggaagg cttctttccc
     4861 atttaggctc agagaacact tttacagaaa tcaaacctat ttacagggtg gcaccccttt
     4921 tctgtggcat ctgatctgtc atctcccaag ctcctgaagt tctttatcca ggttgtatct
     4981 ttgctgagct gacaacagat ttctctatgg atcaaatatt ctaatcaagg aaagatacgg
     5041 tgaaacttca gtcttcttct ggagaa
//
