LOCUS       sequence-403618-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector for human codon optimized SpuFz1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  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 5630)
  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: pMSSM24: pCMV-SpuFz1
FEATURES             Location/Qualifiers
     source          1..5630
                     /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"
     primer_bind     complement(2576..2595)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2641..2696
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2695..2714)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3057..3073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3057..3073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3070..3092)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3081..3097
                     /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(3105..3111,3112..3129,3130..3135))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3150..3171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3230..3425
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3276..3411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3431..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3468..3487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3522..3541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3633..3650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3804..4392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3884..3903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4563..5354,5355..5423))
                     /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     5186..5205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5424..5528)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5559..5630,1..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 tgccgcctaa gaagaagcag aagctggaac ggctgaagaa
      601 actggacaag cccacactgc acacctgtaa caagaccagc ttcgccaagg ccttcctgcc
      661 taacgagaca taccggcagc ggctgctgga ctatatcgcc atcattcacc agctggccga
      721 ccacgcctct cacgccctga agttttacat cctgagcacc tccaccagca gcttccctgt
      781 ggtgcacgag gataccatcg aggccatcct gtacctgctg aacaagggcg aagcctggca
      841 tcccagaaaa gaggccaaga aggcctggcg ggattgcctg ctgccttacg tgcagagata
      901 ctgccagatc gtgggcttca ttcaccccaa cctgagaggc gagcagcaga gcatcaacta
      961 cctgaccgtg tccatgatga ccaacctgaa agtgaacgtg caagagcact tcatgcagat
     1021 gctgctgcgg tacatcaatc tgcgcttcga cgtgaagggc cagaagcaga gactgcctcc
     1081 taagagcgac gcccgcaagg cctttttcac ccggctgaga tacctgaagt ccgtgttcct
     1141 gttcgacgtg gtgcccgagc tggaattcct ggacgatctg acccctctgg aaagcgaggt
     1201 gctggaagag atctggtccc tggacctgcc ttttctgccc aacgatcctc tggcctacgc
     1261 catcgtggcc gatcctatga gcttcttccc agcctactgc aagctgagcg gcctgtacga
     1321 gcagtacggc ttccagcggt ttagcgccat tcctctgcgg agaagcctga tccagtctca
     1381 cgtgcggatc gacaccatca tcctgtatca gcacatcctg tgcatcacca gacgggacgc
     1441 cgagacagtg gaaaaggacg acctgtggat gagagtgtgc aacctgtgca ccaaagcctt
     1501 ccggtccaga tgcggcatgc acttcgaggg cagcatcaca acagatggcg ccagcgtgtc
     1561 cgtgtacctg aaacaccctg aggccgataa gtacggcaag agaggcgcca gaaagtccgc
     1621 caatactgtg gccgccgaag tgaaggccct gtacgtggaa aacaacctgc ctgcctgtag
     1681 agccgccgag aacgtggtgg tcatcgaccc caacaagcgg gatatcctgt actgccagga
     1741 cagcaacggc accaccttca gatacaccgc caaccagaga gccgtggaaa ccggcagcag
     1801 aagattcgcc aagcggcggg aagccatgaa ggaagaagcc ggcgttgacc tgatcgagag
     1861 cagaatccct agccacaaga ccatgaacct gatggacttc accagatacc tgctcgtgcg
     1921 gagagccgac tgggacagac ggaaagagtt ctactctcac cccgctcaca cccggtggaa
     1981 gtggcacagc ttcatcaacc ggcagaagtc cgagagcgac ctgatcagca acatgcggaa
     2041 caaatacggc gagaacttca ccgtggtcat gggcgattgg agcgacgctg gaagaaccgc
     2101 cagattccag accagcagca agaccaaagg ctggcggacc ctgttcaagc ggaaccggat
     2161 cgattgcttc ctgctggacg agtacaagac ctccagcgtg tgccctagat gcagcagcag
     2221 cgagttcgtg gaaaagaagt tcaagacccg gcctcacagc agaccctggc gaagaagaga
     2281 gggcaagatc gagaaggtgc acggcctgct gggctgcacc aatcctaatt gtctgcagca
     2341 ggcctggacc agcggcatga gatactggaa ccgggacatg ctgagcacat gcaacatgct
     2401 gctgatcgtg cggtccatgc tggatggcca tggcagacct gaggtgttct ctagatctgt
     2461 gcctgccgtg gcctgactag cataacccct tggggcgcac tcctcaggtg caggctgcct
     2521 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     2581 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     2641 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     2701 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     2761 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     2821 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     2881 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     2941 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     3001 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     3061 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3121 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3181 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     3241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3421 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3541 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     3601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5521 gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtaa
     5581 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
