LOCUS       sequence-403602-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector for human codon optimized MmeFz2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5150)
  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 5150)
  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: pMJ329: pCMV-MmeFz2
FEATURES             Location/Qualifiers
     source          1..5150
                     /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(2096..2115)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2161..2216
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2215..2234)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2577..2593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2577..2593)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2590..2612)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2601..2617
                     /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(2625..2631,2632..2649,2650..2655))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2670..2691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2750..2945
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2796..2931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2951..3085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2988..3007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3042..3061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3153..3170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3324..3912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3404..3423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4083..4874,4875..4943))
                     /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     4706..4725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4944..5048)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5079..5150,1..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 tgaagcggaa gcgcgagcag atgacactgt ggaaggccgc
      601 cttcgtgaac ggccaagaga cattcaagag ctggatcgac aaggcccgga tgctggaact
      661 gaactgcgac gtgtccagcg ccagcagcac acactacagc gacctgaacc tgaaaacaaa
      721 gtgcgccaag accgacgaca agttcatgtg caactacagc gtgtgcatca gacccaccag
      781 caagcagaag cggaccctga accagatgct gaaggtgtcc aactacgcct acaactggtg
      841 caattacctg gtcaaagaga aggacttcaa gcccaagcag ttcgacctgc agcggatcgt
      901 ggccaagacc aacagcacag atgtgcccgc cgagtacaga ctgcctggcg acgattggtt
      961 cttcgacaac aagatgagca gcatcaagct gaccgcctgc aagaacttct gcaccatgta
     1021 caagagcacc cagaccaacc agaaaaagac caaggtggac ctgcggaaca aggacatcgt
     1081 gcagctgaga gaaggcagct tcgaggtgca gagcaaatac gtgcggctgc tgaccgagaa
     1141 ggacatccct ggcgagagaa tccggcagag cagaatcgcc ctgatgcctg acagcttcag
     1201 caagagcaag aaggactgga aagagcggtt cctgcggctg agcaagaacg tgtccaagat
     1261 tcctccactg agccacgaca tgaaagtgtg caagcggccc aacggcaagt tcatcctgca
     1321 gatcagctgc gaccccatct gcaccagaca gatccaggtg cagaccagcg acagcatctg
     1381 cagcattgat cctggcggca gaaccttcgc cacctgttac gaccccagca acatcaagac
     1441 cttccagatc ggccctgagg ccgacaagaa agagatcatc cacgagttcc acaacaaaat
     1501 cgactacgtg caccggctgc tgtctcacgc ccaagagaag aaacagaccc aggccgtgca
     1561 ggacagaatc ggccagctga agaagctgca tctgaagctc aagacctacg tggacgacgt
     1621 gcacctgaaa ctgtgcagct acctcgtgaa gaactacaag ctggtggtgc tgggcaagat
     1681 cagcgtgtcc tctatcgtgc ggaaggacag acccaaccac ctggccaaga gcgccaacag
     1741 agatctgctg tgctggcagc actaccggtt cagacagagg ctgctgcata gagtgcgggg
     1801 caccgattgc gaagtgatca tccaggacga gcggtacacc tccaagacct gcggcaattg
     1861 cggcgagaag aacaacaaac tcggcggcaa agaaaccttt acctgcgaga gctgcaatta
     1921 caagacccac cgggatgtga acggcgccag aaacatcctg tgcaagtatc tgggactgtt
     1981 ccccttcgcc gcctgactag cataacccct tggggcgcac tcctcaggtg caggctgcct
     2041 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     2101 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     2161 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     2221 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     2281 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     2341 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     2401 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     2461 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     2521 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     2581 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2641 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2701 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     2761 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2821 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2881 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2941 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3001 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3061 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     3121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5041 gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtaa
     5101 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
