LOCUS       sequence-258151-        5042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIRTS-1 (ORF5-TBP6.7-Pin domain-NLS)  in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5042)
  AUTHORS   Rauch S, He E, Srienc M, Zhou H, Zhang Z, Dickinson BC
  TITLE     Programmable RNA-Guided RNA Effector Proteins Built from Human 
            Parts.
  JOURNAL   Cell. 2019 Jun 27;178(1):122-134.e12. doi: 
            10.1016/j.cell.2019.05.049. Epub 2019 Jun 20.
  PUBMED    31230714
REFERENCE   2  (bases 1 to 5042)
  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"; doi: 
            "10.1016/j.cell.2019.05.049"; journalName: "Cell"; date: 
            "2019-06-27- 27"; volume: "178"; issue: "1"; pages: "122-134.e12"
COMMENT     Sequence Label: CIRTS-1: ORF5-TBP6.7-Pin domain-NLS
FEATURES             Location/Qualifiers
     source          1..5042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        230..609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        610..813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     764..784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          949..1081
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1064..1083
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1125..1134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2358..2405
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2412..2435
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2553..2571)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2608..2729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2619..2638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2673..2692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3128..3922
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3182..3201)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3792..3811
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4094..4682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4174..4193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(4798..5042,1..39)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc
       61 cacgatgcgt ccggcgtaga ggatcgagat cttcaatatt ggccattagc catattattc
      121 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
      181 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg gcattgatta
      241 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      301 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      361 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      421 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      481 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      541 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      601 attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca
      661 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      721 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
      781 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcactaga
      841 agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca
      901 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      961 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     1021 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     1081 ggtgtccact cccagttcaa ttacagctct taaggctaga gtatgccacc atggatgacc
     1141 catctttttt gactgggcgc agcacatacg ccaagcgtcg ccgcgcacgc cgtatgaacg
     1201 tctgcaagtg tggtgccatt cttcataaca ataaagactg tcgcagctcc acgatctccg
     1261 gccacaaact tgaccgtttg cgctttgtaa aggaaggtcg tgtagccctt gagggcgaaa
     1321 ctcctgtcta ccgtacctgg gtgaaatggg tcgagaccga atatcacatc aatatcttgg
     1381 agacttcaga tgacgaagag ggcggttctg gaggctcagg ggggagcgga gggtccggtg
     1441 gatcaggagg ctccatggca gttcccgaga cccgccctaa ccacactatt tatatcaaca
     1501 acctcaatag caagatcaag aaggatgagc taaaaaagtc cctgtacgcc atcttctccc
     1561 agtttggcca gatcctggat atcctggtac cgcggcagcg taccccgagg ggccaggcct
     1621 ttgtcatctt caaggaggtc agcagcgcca ccaacgccct gcgctccatg cagggtttcc
     1681 ctttctatga caaacctatg cgtatccagt atgccaagac cgacaaacgt atcccggcca
     1741 agatgaaagg caccttcgtg ggcggtagtg gaggaagcgg ggggagcgga gggagtggtg
     1801 gatccggcgg ctccatggaa ttggagatcc gtccgctttt cctggttccc gataccaatg
     1861 ggttcattga ccacttagcg agcttagccc gtctgcttga gagccgtaag tacattctgg
     1921 tggttcctct tattgtcatc aacgagttag acggccttgc gaaaggccag gaaacagacc
     1981 atcgtgcggg aggctatgcg cgtgtcgtgc aagaaaaggc tcgcaagagc attgagtttt
     2041 tggagcaacg tttcgagtcg cgtgactcct gtttacgtgc acttaccagc cgtggaaatg
     2101 aacttgagtc aatcgcattt cgctctgaag atatcactgg gcagttaggt aacaatgacg
     2161 atttgatttt gtcatgctgc ctgcactatt gtaaagacaa ggcaaaggac tttatgcccg
     2221 cgtcgaagga agagccgatc cgtttgttgc gtgaggtggt attgcttact gatgatcgca
     2281 acctgcgcgt taaggctctt acacgcaacg tccctgtgcg tgacatccca gctttcttaa
     2341 catgggccca ggtcgggaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     2401 aaaagggatc cgattacaag gacgacgatg acaagtagtg tttaaacgaa ttcgggctcg
     2461 gtacacgggg atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     2521 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2581 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2641 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2701 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2761 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     2821 aaatcgaatt ctaatggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     2881 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtgttccgct
     2941 tcctttagca gcccttgcgc cctgagtgct tgcggcagcg tgagcttcaa aagaattgcc
     3001 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     3061 gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgacggtc
     3121 gtttcgcatg cttgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3181 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3241 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3301 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3361 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3421 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3481 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3541 acatcgcatc gagcgagcac gcactcggat ggaagccggt cttgtcgatc aggatgatct
     3601 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgtat
     3661 gccggatggt gaggatctcg tcgtgactca tggcgatgcc tgcttgccga atatcatggt
     3721 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3781 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3841 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3901 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3961 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4021 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4501 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4561 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4621 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     4681 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4741 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     4801 gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg
     4861 cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc
     4921 atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt
     4981 ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga
     5041 tt
//
