LOCUS       sequence-310966-        2391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SV40-AcrIIA4 codon optimized for N.benthamiana.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2391)
  AUTHORS   Calvache C, Vazquez-Vilar M, Selma S, Uranga M, Fernandez-Del-Carmen
            A, Daros JA, Orzaez D
  TITLE     Strong and tunable anti-CRISPR/Cas activities in plants.
  JOURNAL   Plant Biotechnol J. 2022 Feb;20(2):399-408. doi: 10.1111/pbi.13723. 
            Epub 2021 Oct 31.
  PUBMED    34632687
REFERENCE   2  (bases 1 to 2391)
  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: "10.1111/pbi.13723";
            journalName: "Plant Biotechnol J"; date: "2022-02"; volume: "20"; 
            issue: "2"; pages: "399-408"
COMMENT     Sequence Label: pUPD2 SV40-AcrIIA4 (GB3333)
FEATURES             Location/Qualifiers
     source          1..2391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(44..703)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     618..637
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     865..883
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(887..930)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    933..954
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             974..994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1297..1354
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1378..1395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1549..2137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1629..1648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(2320..2391,1..23))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
       61 actcgtcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
      121 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
      181 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
      241 tggtgaaact cacccaggga ttggctgata cgaaaaacat attctcaata aaccctttag
      301 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
      361 gccggaaatc gtcgtggtat tcactccaga gtgatgaaaa cgtttcagtt tgctcatgga
      421 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
      481 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
      541 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
      601 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
      661 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
      721 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
      781 agttggaacc tcttacgtgc ccgatcaact cgagtgccac ctgacgtcta agaaaccatt
      841 attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa
      901 aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agagcgatga
      961 gtggtctcga atgcctaaga aaaaacgaaa ggtcaacatt aatgagctta ttcgtgaaat
     1021 taaaaacaaa gattatacgg ctaagttgag tgggacagac agcaacagta taactcaact
     1081 tatcatacac gtcaataacg acgggaacga gtacgtgatt agcgaatctg aaaacgagtc
     1141 aattgtggag aagttcattt ctgcatttaa gtcaggttgg aatcaggagt atgaagatga
     1201 ggaagaattt tataacgata tgcagaccat aactctgaag tccgaactga attgagcttt
     1261 gagaccactc atcgcgacta gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc
     1321 accaccctgc cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc
     1381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct
     1561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2281 tatatatgag taaacttggt ctgacagctc gaggcttgga ttctcaccaa taaaaaacgc
     2341 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca t
//
