LOCUS       sequence-322049-        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"
     primer_bind     342..361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     595..612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(636..693)
                     /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."
     CDS             complement(996..1016)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1036..1057
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1060..1103
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1107..1125)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1287..1946
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1353..1372)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1967..2061
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(2244..2391,1..441)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
       61 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      421 ggggggcgga gcctgtggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      541 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      601 tcagtgagcg aggaagcctg cataacgcga agtaatcttt tcggttttaa agaaaaaggg
      661 cagggtggtg acaccttgcc cttttttgcc ggactgcagc ggccgctact agtcgcgatg
      721 agtggtctca aagctcaatt cagttcggac ttcagagtta tggtctgcat atcgttataa
      781 aattcttcct catcttcata ctcctgattc caacctgact taaatgcaga aatgaacttc
      841 tccacaattg actcgttttc agattcgcta atcacgtact cgttcccgtc gttattgacg
      901 tgtatgataa gttgagttat actgttgctg tctgtcccac tcaacttagc cgtataatct
      961 ttgtttttaa tttcacgaat aagctcatta atgttgacct ttcgtttttt cttaggcatt
     1021 cgagaccact catcgctcta gaagcggccg cgaattccag aaatcatcct tagcgaaagc
     1081 taaggatttt ttttatctga aattctgcct cgtgatacgc ctatttttat aggttaatgt
     1141 catgataata atggtttctt agacgtcagg tggcactcga gttgatcggg cacgtaagag
     1201 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     1261 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1321 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1381 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     1441 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     1501 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     1561 acaccgtttt ccatgagcaa actgaaacgt tttcatcact ctggagtgaa taccacgacg
     1621 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     1681 cctatttccc taaagggttt attgagaata tgtttttcgt atcagccaat ccctgggtga
     1741 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     1801 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     1861 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     1921 gcgacgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact cctgttgata
     1981 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     2041 gcgtttttta ttggtgagaa tccaagcctc gagctgtcag accaagttta ctcatatata
     2101 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2161 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2221 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2281 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2341 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt t
//
