LOCUS       sequence-449466-        5345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prokaryotic and constitutive expression of AsCas12f-LVA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5345)
  AUTHORS   van der Oost
  TITLE     van der Oost Lab Plasmids 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5345)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pdCas12f1-LVA
FEATURES             Location/Qualifiers
     source          1..5345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(555..1214)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGVA"
     primer_bind     1129..1148
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1215..1317)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1709..1726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1843..2388)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1923..1942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2464..2483
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2578..2612
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     primer_bind     complement(3946..3963)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4116..4202
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4294..4321
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4340..4431
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4889..5344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4976..4995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5186..5207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
       61 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
      121 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
      181 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
      241 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      301 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca
      361 cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc
      421 tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca
      481 ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa
      541 ctgccttaaa aaaattacgc cacgccctgc cactcatcgc agtactgttg taattcatta
      601 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
      661 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
      721 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
      781 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
      841 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
      901 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
      961 catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg
     1021 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     1081 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     1141 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     1201 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1261 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     1321 tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacatggac accaggattt
     1381 atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt
     1441 gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc
     1501 tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag
     1561 caccgccgga catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag
     1621 ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga
     1681 atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc
     1741 gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga
     1801 agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc
     1861 ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact
     1921 ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt
     1981 tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac
     2041 tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt
     2101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa
     2161 aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc
     2221 gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta
     2281 cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt
     2341 ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt
     2401 attaatcaga taaaatattt ctaggctcat gagcccgaag tggcgagccc gatcttcccc
     2461 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgctgg tgatgccggc
     2521 cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcatctg
     2581 acagctagct cagtcctagg tataatgcta gcgagctcag gaggacatca tgatcaaagt
     2641 ttaccgttat gaaatcgtta aaccgctgga cctggattgg aaagaattcg gcaccatcct
     2701 gcgtcagctg cagcaggaaa cccgcttcgc gctgaacaaa gcaacccagc tggcgtggga
     2761 atggatgggt ttcagctctg attacaaaga caaccatggc gaatacccga aaagcaaaga
     2821 catcctgggc tacaccaacg ttcacggtta cgcataccac accatcaaaa ccaaagctta
     2881 tcgtctgaac agcggcaacc tgagccagac catcaaacgt gcgaccgatc gcttcaaagc
     2941 gtaccagaaa gaaatcctgc gtggtgacat gtccatcccg tcttacaaac gtgatatccc
     3001 gctggatctg atcaaagaaa acatcagcgt taaccgtatg aaccacggtg attatatcgc
     3061 ttccctgtcc ctgctgtcta acccggcgaa acaggaaatg aacgttaaac gtaaaatctc
     3121 cgtgattatc atcgttcgtg gcgcgggtaa aaccatcatg gatcgcatcc tgagcggcga
     3181 ataccaggtg agcgcgagcc agatcatcca cgacgaccgt aaaaacaaat ggtacctgaa
     3241 catctcttat gatttcgaac cgcagacccg tgttctggat ctgaacaaaa tcatgggcat
     3301 cgctctgggt gttgcggtgg cggtgtacat ggcgttccag cacaccccgg cgcgctacaa
     3361 actggaaggc ggtgaaatcg aaaacttccg tcgtcaggtg gagtctcgtc gcatctccat
     3421 gctgcgccag ggtaaatacg cgggtggtgc tcgtggcggt cacggccgtg ataaacgtat
     3481 caaaccgatc gaacagctgc gtgacaaaat cgctaacttc cgcgatacca ctaaccaccg
     3541 ttactctcgt tacatcgttg atatggctat caaagaaggt tgcggtacca ttcagatgga
     3601 agatctgacc aacatccgtg atatcggctc tcgtttcctg cagaactgga cctactacga
     3661 tctgcagcag aaaatcatct acaaagcgga agaagcgggc atcaaagtta tcaaaattga
     3721 tccgcagtac accagccagc gttgcagcga atgcggcaac atcgattccg gtaaccgtat
     3781 cggtcaggcg atcttcaaat gccgtgcttg cggctacgaa gcgaacgcgg actacaacgc
     3841 agctcgtaac atcgcgatcc cgaacatcga taaaatcatc gcggaaagca tcaaattagt
     3901 agcttaaaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     3961 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     4021 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     4081 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4141 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4201 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4261 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4321 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4381 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4441 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4501 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc agcaaactat
     4561 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4621 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4681 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4741 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4801 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4861 tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg
     4921 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4981 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5041 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5101 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5161 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5221 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5281 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5341 attta
//
