LOCUS       sequence-325314-        4571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of AcrVIA7 in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4571)
  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 4571)
  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: pAcrVIA7
FEATURES             Location/Qualifiers
     source          1..4571
                     /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(602..619)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      772..858
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      950..977
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        996..1087
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1545..2000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1632..1651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1842..1863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2556..3215)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3130..3149
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3216..3318)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3710..3727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3844..4389)
                     /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(3924..3943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4465..4484
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatgcattaa tacgactcac tataaggagg gctagcaata attttgttta actttaagaa
       61 ggagatatac catgcgcatc attaaactgt atgagcgcat tattcctaag acatcgagca
      121 cttcatacat ctcccgttgg gaggccctga acatccccga tgagaaccgc aatactgcgg
      181 cctggcaccc gcgtacatat cttttttctt acgacaaaga taaagcaatc aatttataca
      241 acacgacaaa tgtcttaggc aattcaggca ttaaaaaacg catcatcgac tacccgtcca
      301 agcgcgaagt ctacatcgcc aatttccctc gcgccatcgc ggacctggta ttgacgatga
      361 aggattacca gttgagttcg ctgcacaact gttgtaatga tttcttcaac gaagatgaga
      421 ccgaacaact ttaccaatac ctgcgcagta tcaaggataa tcgccgcgtc gacgaattct
      481 tgaaatacga attcactgta cgctatttta acgacaaaaa gttttaactc gagcaaagcc
      541 cgccgaaagg cgggcttttc tgtaagcttg gctgttttgg cggatgagag aagattttca
      601 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
      661 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
      721 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
      781 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      841 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
      901 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
      961 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     1021 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1081 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1141 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1201 gggtgcagca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1261 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1321 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1381 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1441 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1501 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
     1561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1741 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1861 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     1921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     1981 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2041 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2221 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2341 cagtcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     2401 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     2461 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     2521 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     2581 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     2641 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     2701 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     2761 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     2821 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     2881 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     2941 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     3001 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     3061 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     3121 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     3181 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     3241 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     3301 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     3361 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     3421 tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt
     3481 ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg
     3541 ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct
     3601 tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg
     3661 ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg
     3721 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     3781 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     3841 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     3961 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     4021 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     4081 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4141 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     4201 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     4261 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     4321 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     4381 cgatctcaag aagatcatct tattaatcag ataaaatatt tctaggctca tgagcccgaa
     4441 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     4501 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     4561 agcttatcat c
//
