LOCUS       sequence-473286-        4802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TaTasR with an N-terminal HA-NLS tag under CMV promoter in
            human cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4802)
  AUTHORS   Faure G, Saito M, Wilkinson ME, Quinones-Olvera N, Xu P, 
            Flam-Shepherd D, Kim S, Reddy N, Zhu S, Evgeniou L, Koonin EV, 
            Macrae RK, Zhang F
  TITLE     TIGR-Tas: A family of modular RNA-guided DNA-targeting systems in 
            prokaryotes and their viruses.
  JOURNAL   Science. 2025 Feb 27:eadv9789. doi: 10.1126/science.adv9789.
  PUBMED    40014690
REFERENCE   2  (bases 1 to 4802)
  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: "Science. 
            2025 Feb 27:eadv9789. doi: 10.1126/science.adv9789."
COMMENT     Sequence Label: pMIIS928
FEATURES             Location/Qualifiers
     source          1..4802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             563..589
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             590..610
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1748..1767)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1813..1868
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1867..1886)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2229..2245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2229..2245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2242..2264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2253..2269
                     /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(2277..2283,2284..2301,2302..2307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2322..2343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2402..2597
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2448..2583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2603..2737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2640..2659)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2694..2713
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2805..2822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2976..3564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3056..3075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3735..4526,4527..4595))
                     /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     4358..4377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4596..4700)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4731..4802,1..304)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atcgccacca tgtaccccta cgacgtgcca gactacgccc ctaagaagaa
      601 gagaaaagtc ggcggctctg gcggcggagg atctggcgga ggtggaagcg gaggagacaa
      661 cgacatcacc atcctggccg tggactggtc ccacgaggaa agaaagctgg ccatcttcga
      721 cggcaagaag atccggaaga agctgcccga gcctagcagc gacgtgatca tcgtggccga
      781 gaacatccct cagaagtacg ccgctccttt catcgaagtg ggcgccaagg tgctgagatg
      841 cagcaccaat gccacagccg acgccagaaa gaactaccag aaaaaggtgg acgccgcctt
      901 cgccaagaac gacgagaacg acagcaaagt gatctgggcc ctgtaccaga cacaccccga
      961 gctgttcaga gagatgaagc tggaacctcc tctgagcagc tactacgcca tcttcaagga
     1021 ctaccaagaa gtgcggatcc ggaccggcaa cagactgtac agcgatagaa ccgacgccat
     1081 ggaagagttc ttcaagattg tgaagaaggg cgagcacgag ctgaagaaag ccgtggacaa
     1141 agagctggaa aatcaccccg tgtacaccca gtggctgcag cacatcaaag gcatcggacc
     1201 tgtggttgcc ggcggactga ttagcctgat cggcgacatc gacagattcg acagcgtgtc
     1261 caagctgtgg gcctacgccg gctactctgt ggacaatggc aaggtgcaga aacggaagaa
     1321 aggcgtcgcc agcaactgga agaacaagat cagaacccac tgctacaaca tcgtggatag
     1381 cttcatcaag cagcggacca gcgtgtacag agagctgtac gacgccgaga aggccagaca
     1441 gaggcctaag gtggaatccg atggacacgc ccacaataga gccgtgcgga aggtggccaa
     1501 agtgttcctg cagcactact gggtcgtgtc tagagagctg gccggcttct ccgtgtctaa
     1561 gccctggatt ctggaacacg gcggccacgt ggactacatc aaacctccac actggaacaa
     1621 ggtggaaatc aagccctgca agccctgact agcataaccc cttggggcgc actcctcagg
     1681 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     1741 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     1801 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     1861 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     1921 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     1981 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     2041 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     2101 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     2161 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     2221 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2281 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2341 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     2401 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2461 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2521 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2581 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     2641 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2701 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     2761 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2881 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2941 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3001 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3061 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3361 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3721 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3781 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3841 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3901 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3961 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4021 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4081 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4141 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4201 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4261 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4321 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4381 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4441 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4501 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4561 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt
     4741 attgactagt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4801 cg
//
