LOCUS       sequence-404728-        5786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector for human codon optimized GtFz1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5786)
  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 5786)
  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: pMJ326: pCMV-GtFz1
FEATURES             Location/Qualifiers
     source          1..5786
                     /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(2732..2751)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2797..2852
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2851..2870)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3213..3229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3213..3229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3226..3248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3237..3253
                     /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(3261..3267,3268..3285,3286..3291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3306..3327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3386..3581
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3432..3567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3587..3721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3624..3643)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3678..3697
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3789..3806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3960..4548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4040..4059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4719..5510,5511..5579))
                     /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     5342..5361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5580..5684)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5715..5786,1..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 tgagcaacat ccggatcgtg aagcggaagg ccaagggctt
      601 cttcaagtgc gaggacctgg tcaccatcaa ggacgccgtg aaagccgctc accggatcat
      661 gtctgatgcc agcattctcg tgcggagcta ctacctgaga tggttccaga gcagctaccc
      721 tctggacagc gacgacaaag agctggaact ggaacacttc cacatcagca tggcctgcag
      781 catcgtgcag ggcatcacaa gacctccagt tagaggcgtg ggccctgagc agagcgtgaa
      841 gatcgacgtg ttcaacgaca tgctggacga gtacaagcgg ctgtacgaga gagcccctaa
      901 cgacaaagaa aacgagacag acctgagcct gagccacgtg ctggcctact ccatcgacaa
      961 tctgctgacc gcctacaaga acaacatcga ggcccacttc agcaaatacg tcaagcggtt
     1021 catccgctgc gatatgctgg ccaagggatt caacaagtct gaggccaaca gagtggccgc
     1081 catctacacc aacgcctaca tctacgacag cagcctggac ctggaacctg acttcatgga
     1141 acggctggga ctcgaggcca ccagctactc tagcctgttt cctagcaaga tcaacaaagg
     1201 cggcttcccc agagtgtacg acctgaaggc taacccctgg gtgtacctgc ctaagatggt
     1261 catgatcaat caggccctgg aaaccgactt ctccagcgtg gaacacaaag agcggcggct
     1321 gctgaaccct ctgcctttct acagcagctt cgtgcccatg cacatcagaa tcgacaccag
     1381 cggcctgtct cagctgctga tgaccaagga cagactggac gacttcaaga gaagctacct
     1441 ggccgagttc ggcgtgtccc tgaacatcaa gaacaaaggg gacatgctcg ccagcttcga
     1501 gaagatcttt ggcagaaagg ccacctccaa tagagaggcc ggactgtacg ctaccgagat
     1561 gtggtccttc ctgaccaacc tgaaaacctg caggcagtgg aaagaactgg acggcgtcgt
     1621 gcggaagaac gaccctaagg gaacccagtg gatgttcgac aacgccgtgg tcacagacgg
     1681 cgtcagcatc agcttccaag tgatcgacaa cagcatgttc ggccgcaagg ccttcagcgg
     1741 ccggaagaaa agagtggcct gtcaagaggc caacgacgaa gaggacagca agcaagtgac
     1801 ccgcgaggaa ctgaaaacca gcaagctgct gggctgcgac cccggcaaga gagatatcct
     1861 ggccatcacc gacggcatca agaccatctg ctacaccaag ggccagagag acatggacac
     1921 ccacaagaca atccggctgc ggaccagcct gaagagaaga agaggttgcg gcctggaaga
     1981 gtacgagaca caagtgatga atcggttcca gaagcggagc tgtcaccccg agatgttcag
     2041 aagatacgcc tgcagccgga agcggatgga acacatgctg ctggaatgct actctcaccc
     2101 cgtgttccgc gagttcaagt tcctggtgta caacaagacc aagagcagcg agcaccggtt
     2161 catgcacaga gtgctggaaa ccttcaagcg gccccagacc aatctgagca aggccagatg
     2221 tgccagcggc gtgatgagaa tgaacgccct gaaagaggtg cagcggcacg gcgatatcat
     2281 catcggctgg ggcaattggg gcaagaaccc caacgctctg agatgctctg ccggacctac
     2341 acctggcatc ggcatcagaa gaagattcga gagcctgttc aagaccacca ccgtgcctga
     2401 gcactacacc agccaagagt gccctagctg caagggcaga tgcctgagaa aagccaccgg
     2461 caatcccatc atgcggcatc atctgctgcg gtgcaccaac gatagctgct gctccagatg
     2521 gtggaaccgg aatgtggctg gcgccttcaa catcctgacc agactgctgg atggccagac
     2581 actgagcggc aatgagacaa caggcgacgg actcggcgga gatgatctgt gactagcata
     2641 accccttggg gcgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     2701 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     2761 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     2821 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     2881 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     2941 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     3001 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     3061 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     3121 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     3181 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3241 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3301 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3361 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     3421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3481 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3541 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     3601 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3661 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     3721 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3781 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3841 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3961 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4021 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4141 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4381 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4561 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4621 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4681 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4741 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4801 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4861 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4921 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4981 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5041 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5101 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5161 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5221 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5281 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5341 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5401 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5461 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5521 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5581 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5641 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5701 gtgccacctg ggtcgacatt gattattgac tagtaatagt aatcaattac ggggtcatta
     5761 gttcatagcc catatatgga gttccg
//
