LOCUS       sequence-205147-        4524 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  T7 promotor drives in vitro transcription of mCherry-tagged mouse 
            TRIM21 with deleted PRYSPRY domain mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4524)
  AUTHORS   Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC, 
            Schuh M
  TITLE     A Method for the Acute and Rapid Degradation of Endogenous Proteins.
  JOURNAL   Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi: 
            10.1016/j.cell.2017.10.033.
  PUBMED    29153837
REFERENCE   2  (bases 1 to 4524)
  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"; journalName: "Cell. 2017 
            Nov 13. pii: S0092-8674(17)31255-2. doi: 
            10.1016/j.cell.2017.10.033."
COMMENT     Sequence Label: pGEMHE-mCherry-mTrim21-Delta-PRYSPRY
FEATURES             Location/Qualifiers
     source          1..4524
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(18..34)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(18..34)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(31..53)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    42..58
                     /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(66..72,73..90,91..96))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    111..132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(249..266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(420..1008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(500..519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1179..1970,1971..2039))
                     /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     1802..1821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2040..2144)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2212..2230
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2268..2290)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2390..2409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2603..2625
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2617..2634
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2618..2634
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2641..2660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2641..2659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2691..2733
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2767..2776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2773..3480
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2918..2936)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3186..3205
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
       61 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      121 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      181 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
      241 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      301 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      361 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      421 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      481 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      541 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      601 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      661 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      721 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      781 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      841 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      901 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      961 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1021 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1081 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     1141 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     1201 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     1261 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     1321 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1381 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1441 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1501 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1561 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1621 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1681 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1741 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1801 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1861 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1921 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1981 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     2041 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2101 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     2161 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     2221 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2281 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2341 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     2401 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2461 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2521 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2581 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
     2641 taatacgact cactataggg cgaattaatt cgagctcggt acccagcttg cttgttcttt
     2701 ttgcagaagc tcagaataaa cgctcaactt tggcagatca attccccggt gctagcgcta
     2761 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2821 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2881 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     2941 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3001 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3061 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3121 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3181 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3241 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3301 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3361 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3421 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3481 tccggactca gatctcgagc tcaagcttcg aattcaatgt ctctggaaaa gatgtgggag
     3541 gaggtcacct gttctatctg cctggatccc atggtggagc ctatgagtat cgaatgtggc
     3601 cattgctttt gcaaggaatg catttttgaa gttgggaaga atgggggcag ttcatgtccc
     3661 gagtgccggc aacagtttct gctccgaaac ctcaggccca atagacatat agccaacatg
     3721 gtggaaaacc ttaaacagat agcccagaat accaagaaga gtacccagga aacgcactgc
     3781 atgaagcatg gagagaagct tcacctattc tgtgaggaag atgggcaggc cctttgctgg
     3841 gtgtgtgccc agtctgggaa acaccgggac cacaccaggg tccctattga agaggctgct
     3901 aaggtatacc aggagaagat ccacgtggct ttagaaaaac tgagaaaggg gaaagagttg
     3961 gccgagaaga tggaaatgga tctcacgatg caaagaacag actggaagag gaacattgac
     4021 acccagaagt cgaggattca cgcagagttc gcacttcaga atagcttgct ggctcaggag
     4081 gagcagaggc agctgcagag gctggagaag gatcaaaggg agtacctgag actcctgggg
     4141 aagaaggagg ctgagctggc tgagaagaac caggccctgc aggagctgat ctcagagctg
     4201 gagaggagga ttcgtggttc agagctggag ctactgcagg aggtgaggat catcctggaa
     4261 aggagtggat cctggaacct ggacacgtta gatattgacg ccccagacct aacaagcaca
     4321 tgccctgtgc cagggcggaa gaagatgctg aggacatgtt gagcggccgc tctagataaa
     4381 tgatagcttg atctggttac cactaaacca gcctcaagaa cacccgaatg gagtctctaa
     4441 gctacataat accaacttac actttacaaa atgttgtccc ccaaaatgta gccattcgta
     4501 tctgctccta ataaaaagaa agtt
//
