LOCUS       sequence-268057-        3681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV and T7 promoter expression plasmid for human codon optimized 
            AcrIIA1 (truncated C-terminal domain only) with C-terminal NLS 
            (SV40).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3681)
  AUTHORS   Osuna BA, Karambelkar S, Mahendra C, Christie KA, Garcia B, Davidson
            AR, Kleinstiver BP, Kilcher S, Bondy-Denomy J
  TITLE     Listeria Phages Induce Cas9 Degradation to Protect Lysogenic 
            Genomes.
  JOURNAL   Cell Host Microbe. 2020 Apr 18. pii: S1931-3128(20)30190-6. doi: 
            10.1016/j.chom.2020.04.001.
  PUBMED    32325050
REFERENCE   2  (bases 1 to 3681)
  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: "Cell Host 
            Microbe. 2020 Apr 18. pii: S1931-3128(20)30190-6. doi: 
            10.1016/j.chom.2020.04.001."
COMMENT     Sequence Label: pCMV-T7-AcrIIA1(CTD)-NLS(SV40) (KAC438)
FEATURES             Location/Qualifiers
     source          1..3681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             916..936
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             945..962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(985..1002)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    991..1215
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1286..1302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1286..1302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1299..1321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1310..1326
                     /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(1334..1340,1341..1358,1359..1364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1379..1400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1517..1534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1688..2276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1768..1787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2447..3238,3239..3307))
                     /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     3070..3089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3308..3412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3491..3510)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 gacgacctgg ccggcttcaa gcacctgctg gacaagtaca agctgagctt ccccgcccag
      721 gagttcgagc tgtactgcct gatcaaggag ttcgagagcg ccaacatcga ggtgctgccc
      781 ttcaccttca acagattcga gaacgaggag cacgtgaaca tcaagaagga cgtgtgcaag
      841 gccctggaga acgccatcac cgtgctgaag gagaagaaga acgagctgct gggatccggc
      901 ggtggaggca gtgggcccaa gaagaagagg aaagtctgac cggtcatcat caccatcacc
      961 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1021 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1081 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1141 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1201 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgata ccgtcgacct
     1261 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1321 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctag ggtgcctaat
     1381 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1441 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     1501 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1561 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1621 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2101 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2221 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2401 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2581 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3301 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3421 cccgaaaagt gccacctgac gtcgacggat cgggagatcg atctcccgat cccctagggt
     3481 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     3541 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     3601 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     3661 tacgggccag atatacgcgt t
//
