LOCUS       sequence-220499-        3960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-T7 promoter expression plasmid for human codon optimized AcrVA1 
            anti-CRISPR protein with C-terminal NLS .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3960)
  AUTHORS   Marino ND, Zhang JY, Borges AL, Sousa AA, Leon LM, Rauch BJ, Walton 
            RT, Berry JD, Joung JK, Kleinstiver BP, Bondy-Denomy J
  TITLE     Discovery of widespread Type I and Type V CRISPR-Cas inhibitors.
  JOURNAL   Science. 2018 Sep 6. pii: science.aau5174. doi: 
            10.1126/science.aau5174.
  PUBMED    30190308
REFERENCE   2  (bases 1 to 3960)
  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: "Science. 
            2018 Sep 6. pii: science.aau5174. doi: 10.1126/science.aau5174."
COMMENT     Sequence Label: pCMV-T7-hAcrVA1-NLS(sv40) (BPK5050)
FEATURES             Location/Qualifiers
     source          1..3960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             538..558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             567..584
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(607..624)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    613..837
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(908..924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(908..924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(921..943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    932..948
                     /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(956..962,963..980,981..986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1001..1022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1139..1156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1310..1898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1390..1409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2069..2860,2861..2929))
                     /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     2692..2711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2930..3034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3113..3132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3304..3683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3684..3887
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3838..3858
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3929..3948
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3929..3947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(3955..3960,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggctatg aggcaaaaga acggtacgct aaaaagaaga tgcaggagaa tacaaaaata
       61 gacacactta cagacgagca gcacgacgca ctggcccaac tgtgtgcctt ccgacataaa
      121 tttcactcca ataaggacag tctcttcctc tctgagtccg cttttagcgg tgagttttcc
      181 tttgagatgc aaagcgatga aaatagcaag ctgagagaag taggtcttcc cacaatagaa
      241 tggtcattct acgacaacag tcacattccg gacgatagct tccgagagtg gtttaacttt
      301 gcaaactact cagagctctc tgagacaata caagaacagg gtctggagct ggacctcgac
      361 gacgatgaga catacgagct ggtttatgat gaactctata cagaggctat gggagaatat
      421 gaagagctta atcaagatat tgagaagtat ctgaggagga ttgacgagga acatgggacc
      481 cagtattgcc caaccgggtt tgcccgcctt cgaggatccg gcggtggagg cagtgggccc
      541 aagaagaaga ggaaagtctg accggtcatc atcaccatca ccattgagtt taaacccgct
      601 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
      661 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
      721 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
      781 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
      841 ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag agcttggcgt
      901 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      961 tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc taactcacat
     1021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2041 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2101 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2161 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2221 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2281 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2341 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2401 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2461 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2521 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2581 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2641 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2701 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2761 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2821 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2881 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2941 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3001 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3061 acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc agtacaatct
     3121 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     3181 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     3241 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     3301 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3361 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3421 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3481 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3541 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3601 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     3661 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     3721 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     3781 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     3841 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     3901 gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag agccgccacc
//
