LOCUS       sequence-325315-        4346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of AcrVIA4 in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4346)
  AUTHORS   Wandera KG, Beisel CL
  TITLE     Rapidly Characterizing CRISPR-Cas13 Nucleases Using Cell-Free 
            Transcription-Translation Systems.
  JOURNAL   Methods Mol Biol. 2022;2404:135-153. doi: 
            10.1007/978-1-0716-1851-6_7.
  PUBMED    34694607
REFERENCE   2  (bases 1 to 4346)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2404:135-153. doi"; pages: " 
            10.1007/978-1-0716-1851-6_7"
COMMENT     Sequence Label: pAcrVIA4
FEATURES             Location/Qualifiers
     source          1..4346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             43..65
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(377..394)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      547..633
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      725..752
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        771..862
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1320..1775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1407..1426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1617..1638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2331..2990)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2905..2924
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2991..3093)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3485..3502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3619..4164)
                     /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(3699..3718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4240..4259
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatgcattaa tacgactcac tataaggagg gctagcaata attttgttta actttaagaa
       61 ggagatatac catggataaa gcgaaccgct gccttaaagc gaaggacaaa attctgaata
      121 ttcttgaaaa agaagaaatt actcttgacg aatttaataa catcagcaag gatatcgcaa
      181 aagaatacgt tgaaaaagca gttctgaagc ccaaggatat tgcagagcgt atcatcaaca
      241 tggtcaaaaa cgctaagtct atcagtttcg atgagctggc atcagaaatc tccgaagagt
      301 aactcgagca aagcccgccg aaaggcgggc ttttctgtaa gcttggctgt tttggcggat
      361 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
      421 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
      481 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
      541 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      601 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
      661 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
      721 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
      781 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      841 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      901 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      961 tgctgaagat cagttgggtg cagcaaacta ttaactggcg aactacttac tctagcttcc
     1021 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1081 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1141 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1201 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1261 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1501 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     1561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1621 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     1681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1741 cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt
     1801 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1861 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1921 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1981 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2041 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2101 tgctaatcct gttaccagtc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     2161 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     2221 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt
     2281 caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg
     2341 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     2401 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     2461 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     2521 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     2581 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     2641 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     2701 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     2761 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     2821 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     2881 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     2941 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     3001 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     3061 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc
     3121 cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc
     3181 gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt actgatttag
     3241 tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg
     3301 ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctagcg
     3361 gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg
     3421 gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg
     3481 cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt
     3541 acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag
     3601 ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg
     3661 acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3721 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     3781 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     3841 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     3901 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     3961 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     4021 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     4081 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     4141 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
     4201 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     4261 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     4321 ctgctcatgt ttgacagctt atcatc
//
