LOCUS       sequence-249186-        4202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AcrX expression in mammalian cells; AcrX is an engineered 
            anti-CRISPR protein targeting S. aureus Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4202)
  AUTHORS   Mathony J, Harteveld Z, Schmelas C, Upmeier Zu Belzen J, 
            Aschenbrenner S, Sun W, Hoffmann MD, Stengl C, Scheck A, Georgeon S,
            Rosset S, Wang Y, Grimm D, Eils R, Correia BE, Niopek D
  TITLE     Computational design of anti-CRISPR proteins with improved 
            inhibition potency.
  JOURNAL   Nat Chem Biol. 2020 Apr 13. pii: 10.1038/s41589-020-0518-9. doi: 
            10.1038/s41589-020-0518-9.
  PUBMED    32284602
REFERENCE   2  (bases 1 to 4202)
  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: "Nat Chem 
            Biol. 2020 Apr 13. pii: 10.1038/s41589-020-0518-9. doi: 
            10.1038/s41589-020-0518-9."
COMMENT     Sequence Label: AcrIIC1X
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    989..1110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1026..1045)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1080..1099
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1117..1572)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1254..1275)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1466..1485
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1599..1703
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1705..2062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1705..1896
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1728..1748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1913..2048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2097..2891
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2097..2891
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2151..2170)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2761..2780
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3079..3098)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3123..3170
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3499..4087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3988..4007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgccac
      601 catggcattc aaaacgtaca agataggcaa aaacgctggg taccaggggt gtggtctgtg
      661 cctcgctgcg atcagcgaga atgaggctat caaggtcaaa tacttgcgcg acatctgccc
      721 ggactacgat ggcgacgaca aaatcgaaga ttggttgagg tgggggacgg attctcgcgt
      781 taaggccgct gcgcttgaaa tggagcaata tgcctacaca tccgtcggta tggcgtcctg
      841 ttgggagttc gttgagttgt gactcgagcg gccgcgactc tagatcataa tcagccatac
      901 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
      961 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1021 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1081 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1141 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1201 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1261 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1321 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1381 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1441 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1501 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1561 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1621 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1681 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     1741 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1801 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1861 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     1921 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     1981 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2041 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2101 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2161 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2221 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2281 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2341 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2401 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2461 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2521 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2581 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2641 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2701 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2761 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2821 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2881 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     2941 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3001 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3061 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3121 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3181 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3241 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3301 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3361 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3601 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4081 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4141 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     4201 tt
//
