LOCUS       sequence-224368-        8637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AcrVA1 in human cells, contained in a lentiviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8637)
  AUTHORS   Watters KE, Fellmann C, Bai HB, Ren SM, Doudna JA
  TITLE     Systematic discovery of natural CRISPR-Cas12a inhibitors.
  JOURNAL   Science. 2018 Oct 12;362(6411):236-239. doi: 
            10.1126/science.aau5138. Epub 2018 Sep 6.
  PUBMED    30190307
REFERENCE   2  (bases 1 to 8637)
  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"; doi: 
            "10.1126/science.aau5138"; journalName: "Science"; date: 
            "2018-10-12- 12"; volume: "362"; issue: "6411"; pages: "236-239"
COMMENT     Sequence Label: pCF525-EF1a-Hygro-P2A-AcrVA1-lenti
FEATURES             Location/Qualifiers
     source          1..8637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             6..186
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    233..358
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    851..1084
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1269..1313
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1611..1728
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1803..2980
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2034..2971
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2928..2948
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2998..3007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3004..4029
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MAKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             4039..4095
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    4636..5224
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4689..4709)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5107..5118)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5296..5529
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5555..5572)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5561..5785
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5891..5908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6062..6650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6142..6161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6821..7612,7613..7681))
                     /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     7444..7463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7682..7786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7861..7880)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8046..8425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8427..8625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8580..8600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
       61 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      121 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      181 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
      241 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
      301 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
      361 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
      421 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
      481 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
      541 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
      601 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
      661 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
      721 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
      781 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
      841 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
      901 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
      961 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     1021 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     1081 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     1141 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     1201 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     1261 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     1321 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     1381 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     1441 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     1501 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     1561 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     1621 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     1681 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     1741 agggacagca gagatccagt ttggttaatt aagctagcta ggtcttgaaa ggagtgggaa
     1801 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     1861 ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa
     1921 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     1981 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2041 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2101 ttacttccac tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     2161 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     2221 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     2281 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     2341 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     2401 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     2461 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     2521 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     2581 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     2641 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     2701 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     2761 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     2821 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     2881 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     2941 agtggttcaa agtttttttc ttccatttca ggtgtcgtga accggttcta gagcgctgcc
     3001 accatggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     3061 ttcgacagcg tgtccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     3121 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     3181 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     3241 gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     3301 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggca
     3361 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     3421 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     3481 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     3541 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     3601 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     3661 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     3721 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     3781 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     3841 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     3901 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     3961 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     4021 gcaaaggaag gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa
     4081 gagaatcctg gaccgatgtc aaaggctatg tatgaagcca aagaacgata tgcgaagaag
     4141 aagatgcagg agaacactaa gatagacact ctcaccgatg agcagcatga cgctctcgcc
     4201 cagctttgcg cgtttagaca taagtttcat tctaacaaag attccctgtt tctttccgaa
     4261 tcagcgttca gtggggagtt tagtttcgag atgcagtctg acgagaactc caagttgagg
     4321 gaagtaggac tccccacaat cgaatggtcc ttttacgaca attctcacat acccgacgac
     4381 agctttagag agtggttcaa ttttgctaac tattctgagc tttcagaaac aatacaagag
     4441 cagggtcttg aactcgacct ggatgatgac gaaacctacg agctggtcta tgatgagctt
     4501 tatacagagg cgatgggaga atacgaggaa ttgaaccagg atatagagaa ataccttcgc
     4561 agaatagatg aggaacatgg cacgcaatac tgtccaactg gttttgcgcg gctgagataa
     4621 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4681 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4741 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4801 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4861 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4921 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4981 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5041 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5101 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5161 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5221 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     5281 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     5341 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5401 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5461 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5521 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     5581 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     5641 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     5701 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     5761 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagcca ttaatgaaaa
     5821 gaaccagcca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5881 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7801 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     7861 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7921 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     7981 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     8041 atattgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     8101 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     8161 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     8221 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     8281 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     8341 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     8401 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     8461 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     8521 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     8581 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcct
//
