LOCUS       sequence-275297-        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"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             667..687
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             696..713
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(736..753)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    742..966
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1037..1053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1037..1053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1050..1072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1061..1077
                     /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(1085..1091,1092..1109,1110..1115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1130..1151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1268..1285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1439..2027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1519..1538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2198..2989,2990..3058))
                     /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     2821..2840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3059..3163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3242..3261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(3433..3681,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggacgacctg
      421 gccggcttca agcacctgct ggacaagtac aagctgagct tccccgccca ggagttcgag
      481 ctgtactgcc tgatcaagga gttcgagagc gccaacatcg aggtgctgcc cttcaccttc
      541 aacagattcg agaacgagga gcacgtgaac atcaagaagg acgtgtgcaa ggccctggag
      601 aacgccatca ccgtgctgaa ggagaagaag aacgagctgc tgggatccgg cggtggaggc
      661 agtgggccca agaagaagag gaaagtctga ccggtcatca tcaccatcac cattgagttt
      721 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
      781 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
      841 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
      901 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
      961 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
     1021 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1081 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
     1141 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1621 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1861 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3181 tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg tcgactctca
     3241 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     3301 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     3361 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     3421 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     3481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     3541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     3601 cgccaatagg gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact
     3661 tggcagtaca tcaagtgtat c
//
