LOCUS       sequence-403503-        3040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector for SpuFz1 omegaRNA 75-nt scaffold + 5' MS2-
            (BbsI guide entry site).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3040)
  AUTHORS   Saito M, Xu P, Faure G, Maguire S, Kannan S, Altae-Tran H, Vo S, 
            Desimone A, Macrae RK, Zhang F
  TITLE     Fanzor is a eukaryotic programmable RNA-guided endonuclease.
  JOURNAL   Nature. 2023 Jun 28. doi: 10.1038/s41586-023-06356-2.
  PUBMED    37380027
REFERENCE   2  (bases 1 to 3040)
  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: "Nature. 
            2023 Jun 28. doi: 10.1038/s41586-023-06356-2."
COMMENT     Sequence Label: pMJ548: pU6-SpuFz1_MS2-omegaRNA(75nt)-entry
FEATURES             Location/Qualifiers
     source          1..3040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..29)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        14..343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      194..329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     256..275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    349..470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(386..405)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     440..459
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(519..535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(519..535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(532..554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    543..559
                     /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(567..573,574..591,592..597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(750..767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(921..1509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1001..1020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1680..2471,2472..2540))
                     /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     2303..2322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2541..2645)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2671..2911
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     2671..2691
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2842..2861
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2917..2935
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
       61 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
      181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
      241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
      301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
      361 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      421 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
      481 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
      541 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      601 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      901 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1021 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1081 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1141 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1201 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1261 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1321 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1381 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1441 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1501 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1561 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1621 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1681 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1741 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1801 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1861 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1921 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1981 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2041 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2101 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2161 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2221 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2281 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2341 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2401 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2521 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2641 ccgcgcacat ttccccgaaa agtgccacct gagggcctat ttcccatgat tccttcatat
     2701 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2761 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2821 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2881 ttcttggctt tatatatctt gtggaaagga cagaaaacat gaggatcacc catgtctgca
     2941 gttttccgag ccggttgtcg cgcggttcaa tccctggtgc gggtgctagt gccaataccc
     3001 accggctccg cactacagtc ttctctgaag accatttttt
//
