LOCUS       sequence-449468-        5939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prokaryotic and constitutive expression of AsCas12f1/TadA8e-LVA 
            fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5939)
  AUTHORS   van der Oost
  TITLE     van der Oost Lab Plasmids 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5939)
  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            "Unpublished"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: pdCas12f1Cas_ABE-LVA
FEATURES             Location/Qualifiers
     source          1..5939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..84
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(151..253)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(645..662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(779..1324)
                     /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(859..878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1400..1419
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1514..1548
                     /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(3476..3493)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3646..3732
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3824..3851
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3870..3961
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4419..4874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4506..4525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4716..4737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5430..5939,1..150))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
       61 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
      121 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
      181 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
      241 tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
      301 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
      361 gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag
      421 gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg
      481 tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta
      541 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
      601 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
      661 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
      721 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
      781 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
      841 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
      901 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
      961 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     1021 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1081 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     1141 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     1201 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     1261 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     1321 tcaagaagat catcttatta atcagataaa atatttctag gctcatgagc ccgaagtggc
     1381 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     1441 cgctggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
     1501 atcatcgatg catctgacag ctagctcagt cctaggtata atgctagcga gctcaggagg
     1561 acatcatgat caaagtttac cgttatgaaa tcgttaaacc gctggacctg gattggaaag
     1621 aattcggcac catcctgcgt cagctgcagc aggaaacccg cttcgcgctg aacaaagcaa
     1681 cccagctggc gtgggaatgg atgggtttca gctctgatta caaagacaac catggcgaat
     1741 acccgaaaag caaagacatc ctgggctaca ccaacgttca cggttacgca taccacacca
     1801 tcaaaaccaa agcttatcgt ctgaacagcg gcaacctgag ccagaccatc aaacgtgcga
     1861 ccgatcgctt caaagcgtac cagaaagaaa tcctgcgtgg tgacatgtcc atcccgtctt
     1921 acaaacgtga tatcccgctg gatctgatca aagaaaacat cagcgttaac cgtatgaacc
     1981 acggtgatta tatcgcttcc ctgtccctgc tgtctaaccc ggcgaaacag gaaatgaacg
     2041 ttaaacgtaa aatctccgtg attatcatcg ttcgtggcgc gggtaaaacc atcatggatc
     2101 gcatcctgag cggcgaatac caggtgagcg cgagccagat catccacgac gaccgtaaaa
     2161 acaaatggta cctgaacatc tcttatgatt tcgaaccgca gacccgtgtt ctggatctga
     2221 acaaaatcat gggcatcgct ctgggtgttg cggtggcggt gtacatggcg ttccagcaca
     2281 ccccggcgcg ctacaaactg gaaggcggtg aaatcgaaaa cttccgtcgt caggtggagt
     2341 ctcgtcgcat ctccatgctg cgccagggta aatacgcggg tggtgctcgt ggcggtcacg
     2401 gccgtgataa acgtatcaaa ccgatcgaac agctgcgtga caaaatcgct aacttccgcg
     2461 ataccactaa ccaccgttac tctcgttaca tcgttgatat ggctatcaaa gaaggttgcg
     2521 gtaccattca gatggaagat ctgaccaaca tccgtgatat cggctctcgt ttcctgcaga
     2581 actggaccta ctacgatctg cagcagaaaa tcatctacaa agcggaagaa gcgggcatca
     2641 aagttatcaa aattgatccg cagtacacca gccagcgttg cagcgaatgc ggcaacatcg
     2701 attccggtaa ccgtatcggt caggcgatct tcaaatgccg tgcttgcggc tacgaagcga
     2761 acgcggacta caacgcagct cgtaacatcg cgatcccgaa catcgataaa atcatcgcgg
     2821 aaagcatcaa atctggtggt tcttctggtg gttctagcgg cagcgagact cccgggacct
     2881 cagagtccgc cacacccgaa agttctggtg gttcttctgg tggttcttct gaggtggagt
     2941 tttcccacga gtactggatg agacatgccc tgaccctggc caagagggca cgggatgaga
     3001 gggaggtgcc tgtgggagcc gtgctggtgc tgaacaatag agtgatcggc gagggctgga
     3061 acagagccat cggcctgcac gacccaacag cccatgccga aattatggcc ctgagacagg
     3121 gcggcctggt catgcagaac tacagactga ttgacgccac cctgtacgtg acattcgagc
     3181 cttgcgtgat gtgcgccggc gccatgatcc actctaggat cggccgcgtg gtgtttggcg
     3241 tgaggaactc aaaaagaggc gccgcaggct ccctgatgaa cgtgctgaac taccccggca
     3301 tgaatcaccg cgtcgaaatt accgagggaa tcctggcaga tgaatgtgcc gccctgctgt
     3361 gcgatttcta tcggatgcct agacaggtgt tcaatgctca gaagaaggcc cagagctcca
     3421 tcaacttagt agcttaaaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     3481 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     3541 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     3601 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     3661 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     3721 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     3781 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     3841 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
     3901 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3961 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4021 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4081 agcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4141 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4201 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4261 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4321 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4381 tcagaccaag tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt
     4441 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4501 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4561 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4621 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4681 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4741 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     4801 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta ataaaatatt
     4861 aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5101 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtca
     5221 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     5281 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct
     5341 ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt
     5401 taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac
     5461 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     5521 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     5581 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     5641 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     5701 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     5761 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     5821 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     5881 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttatt
//
