LOCUS       sequence-275298-        3915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV and T7 promoter expression plasmid for human codon optimized 
            Leu-AcrIIA1 with C-terminal NLS (SV40).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3915)
  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 3915)
  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-Leu-NLS(SV40) (KAC440)
FEATURES             Location/Qualifiers
     source          1..3915
                     /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             901..921
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             930..947
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(970..987)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    976..1200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1271..1287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1271..1287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1284..1306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1295..1311
                     /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(1319..1325,1326..1343,1344..1349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1364..1385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1502..1519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1673..2261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1753..1772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2432..3223,3224..3292))
                     /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     3055..3074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3293..3397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3476..3495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(3667..3915,1..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 gggcaagctg
      421 gacgactacc tgaagctgaa caacaccacc agatacgagg tggccaagat cagcggcatc
      481 cccgagacca gcttcaagag catcagaaac agagacgtga acaacctgag cggcagattc
      541 tacagagcca tcggcctggt gctgggcaag accggcggcc aggtgtacga cgagatcacc
      601 gccgacgaga acaccgtgtt caacttcctg ggcaagcacc acatccacga caaggagaga
      661 gtgaccgagc tgctggacta catgctgtac ttcaagaagc acgacatcga cgtgaccaac
      721 gtgagcttca acagattcga gaacgagatc gagaacggcc acatcctggg cgacgaggac
      781 gacgtgctgc aggtgatcga caacctgatc gagagcttca agaccatgaa ggagaacgtg
      841 gaggccggca acctgcccac ccccgagaag atggacggat ccggcggtgg aggcagtggg
      901 cccaagaaga agaggaaagt ctgaccggtc atcatcacca tcaccattga gtttaaaccc
      961 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1021 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1081 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1141 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     1201 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     1261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1321 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     1381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1861 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2101 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3421 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
     3481 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     3541 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     3601 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     3661 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     3721 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3781 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3841 tagggacttt ccattgacgt caatgggtgg actatttacg gtaaactgcc cacttggcag
     3901 tacatcaagt gtatc
//
