LOCUS       sequence-471745-        4823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ParTasR 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 4823)
  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 4823)
  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: pMIIS1011
FEATURES             Location/Qualifiers
     source          1..4823
                     /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(1769..1788)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1834..1889
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1888..1907)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2250..2266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2250..2266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2263..2285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2274..2290
                     /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(2298..2304,2305..2322,2323..2328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2343..2364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2423..2618
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2469..2604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2624..2758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2661..2680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2715..2734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2826..2843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2997..3585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3077..3096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3756..4547,4548..4616))
                     /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     4379..4398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4617..4721)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4752..4823,1..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 gaggaagaca
      661 ggtggccatc gattgggcct tcgccgaaga aaagctggcc gtgtacgacg gcaagaaagt
      721 gctgaaggcc gtgcctaagc tgaaagccgg cgacgagatc ttcgccgaga acatccctat
      781 gaagcacgcc gctcagtggc tgaaggacgg catcatcatc agacggtgca gacccaacga
      841 taccgccgct ctgagaaaag agatcggcca agagaaaaca gacgccctgg acgccaagct
      901 gatctggcaa ctggctgagg cccatcctga gaagttcaga gagtggaagg gcgaccctca
      961 gctgaccaca ctgtaccggg ccttcaaaga ggtgcagcgc tgtagagtgg gccagagcaa
     1021 tagagtgtgg gccaagggcg aagagacagc cacagcagtg ctggacgacc tggaaaagac
     1081 cgagcggaag atcgtgaagg ccatcgagaa agagctgaaa agctacaaag tgtgggactg
     1141 gctgtcccag atcaaaggca tcggagtggc tacaggcgga ggactcgttg gcctgatcgc
     1201 caaatacggc atcgagaata tcagacaggt gtccagcctg tggcacctgt ttggactgca
     1261 tgtggtggaa ggcaaggccc ctagacggac aaaaggcgaa gaggtgtcct acgtgcccga
     1321 ggccaaaaca ctggtgctgg gagtgatcgc cgactgcttc atcaagcaga gaagccccgt
     1381 gtaccgggac atctacgacc aagagaaggc cagacagctg gaaatcgagt accctgaggg
     1441 cgagctggcc tccaagttcc tgggctacaa gaagtctgac acccacctga gcctgaatca
     1501 cgcccacaga cgcgccatcc ggaagatgat gaagatcttt gtgcagcacg tgtggctggc
     1561 ctggcggaca tgtgaaggcc tggaaaccag acctctgtac tgccacgagt acctgggcca
     1621 cgagcacttc attgagcctc cagtgaagat ccagccagag ggctcttgac tagcataacc
     1681 ccttggggcg cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     1741 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     1801 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     1861 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1921 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     1981 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     2041 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     2101 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     2161 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     2221 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2281 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2341 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2401 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     2461 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2521 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2581 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     2641 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2701 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     2761 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2821 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2881 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2941 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3001 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3061 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3121 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3181 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3241 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3301 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3361 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3421 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3481 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3541 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3601 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3661 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3721 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3781 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3841 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3901 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3961 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4021 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4081 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4141 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4201 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4261 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4321 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4381 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4441 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4501 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4561 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4621 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4681 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4741 ccacctgggt cgacattgat tattgactag taatagtaat caattacggg gtcattagtt
     4801 catagcccat atatggagtt ccg
//
