LOCUS       Cas9d_pET28a            7929 bp    DNA     circular     13-MAY-2025
DEFINITION  34-1 in pET-28a(+).
ACCESSION   urn.local...2rn-j5vvk8s
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     Resistance markers: Kanamycin                                      
                                                                               
                                         This file was created using tools
            provided by Twist Bioscience.
FEATURES             Location/Qualifiers
     source          1..7929
                     /organism="34-1 in pET-28a(+)"
     misc_feature    337..2583
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: Effector"
                     /standard_name="34-1"
     misc_feature    337..357
                     /note="Geneious type: overhang"
                     /standard_name="start"
     misc_feature    2784..3090
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: rep_origin"
                     /standard_name="f1"
     misc_feature    3314..4129
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: Antibiotic R"
                     /standard_name="kan"
     misc_feature    4251..4839
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: rep_origin"
                     /standard_name="colE1_high_copy"
     misc_feature    5266..5457
                     /note="Geneious type: misc_feature_"
                     /standard_name="ROP"
     misc_feature    6232..6253
                     /note="Geneious type: misc_feature_"
                     /standard_name="CAP_binding"
     misc_feature    6266..7357
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: ROI"
                     /standard_name="lacI"
     misc_feature    7349..7426
                     /note="Geneious type: misc_feature_"
                     /standard_name="lacIQ_promoter"
     misc_feature    7735..7753
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: promoter"
                     /standard_name="T7"
     misc_feature    7753..7780
                     /note="Geneious type: misc_feature_"
                     /standard_name="lacO"
     misc_feature    7793..7815
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: RBS"
                     /standard_name="RBS_3"
     misc_feature    7834..7850
                     /note="Geneious type: misc_feature_"
                     /standard_name="6x_His_tag"
ORIGIN      
        1 atgagcgatt ccgaagttaa tcaagaagcg aaacctgagg tgaaaccgga ggtgaaaccg
       61 gaaacacata ttaatttaaa agtctctgat gggagctctg aaattttttt taaaatcaaa
      121 aaaacaacac ctcttcggag attgatggaa gcctttgcaa aacgccaagg caaagaaatg
      181 gattcactgc gctttctgta cgacggtatt cgtattcagg cggaccagac accagaggat
      241 ctggacatgg aagataatga tatcattgaa gcccatcgcg aacaaattgg ggagaactta
      301 tattttcagg gtcacatgat ggcagcggct ttctcaatgg agagagaact ggtcttgggt
      361 atcgactacg gtggaaagta tacaggatta gcagtggttg atagacgtca taatcaggtt
      421 ctgtatgcaa accgcctcaa aatgagagat gatgtcgctg gtatcctgaa agaccgccgg
      481 aagcaacgag ggattcgccg tacagcgcag acaaaaaaaa aacgcctgcg ggaactgaag
      541 aattatttga aaagtatagg ttacaacgaa agtaccgcaa cctttgagac ggtctatagc
      601 ttagcccata aacgaggata cgattatgcg gatatgcctg aagaaaagac ctcggaggaa
      661 atagaagcaa tggacgtgga agagcgtaag cagtgggaaa aagaaaagca ggaatgggaa
      721 gaaacaaagc gcaattcccg ccatcggaaa gaagtggtga aagatgtgca taaggcaatg
      781 atagaaggcc gtgcaactga ggaacagatt aagagagtgg aacgcatatt caacaaacag
      841 tacagaccca aacgtttcaa taatcgcata cttaccaaat gcaaagtgga agattgtggg
      901 gtcaatacac ctctgcgtaa gaacgtgcgt gatctgttaa tcgaaaatat tgtgcgtttc
      961 ttcccgattg aacagtcgga aaaggacaat ctgaaagacg cagtcctgga taaaaatcgc
     1021 cgggaggaag tgaaaagttt ttttcgcaaa cataaaactg acgaacacat tcgcaaacaa
     1081 gtgtatgata tcgcggacaa taaattgagt gggcgtaccg ttttctgcaa ggaacatatt
     1141 ctcgagcata cagaacactc gaaggaagaa agaaaagtat ttcggctcgc cccttctctc
     1201 aaaactaaaa ttgaaaatgt tttagcagta attaaagatg agatattacc aaaattcacg
     1261 gtaaacaagg ttgttatgga gtcgaacaac tttgatattg cagccaaaac ccagggcaag
     1321 aaaagattag caaaagaaga gtatggcaaa ggtcctcgcg aaggcaagga gacccgcaag
     1381 gaagcactgc ttcgtgaaac agatgggcgt tgtatttatt gcggaaagag tatcgatatt
     1441 tctaatgccc acgatgacca tatttttcct agaaaagcag gagggctgaa catctttgcg
     1501 aacctggtgg cttgctgcgc ggtctgtaat gaaaataaaa aggggcgcac gccactggaa
     1561 tcgggcatct ctcctaaacc tgaaattatt gcgtttatga agaatgatct taaaaaaaaa
     1621 attctggagg acgcgcggaa tataaatacg gtggacttta ataaatacat gtctcacgca
     1681 tcaatcgggt ggcgctatat gcgggatcgt cttcgtgaat cagcaggtaa taagaaactc
     1741 ccgatagaac gccagtcggg tatctatacg gcctattttc gtagatggtg ggggttcaaa
     1801 aaagaacgtg gcaacacgct gcaccacgca cttgacgcag tgattttagc atcacggaaa
     1861 ggctattccg atgatggttt agtggatatg acgcttaaac ctaagtataa caaaggtgga
     1921 gaatttgatc cggaaaagca cttacccgag cctatcgagt ttaagatgga taagggtagt
     1981 agaggttctg ccttgcacga tcgtaatcca ttatcgtata aaaaaggcat cattacacgg
     2041 cgtttcatgg ttactgaaat cgagtgtgga aaagaagacg atgtgatttc cgaaacttat
     2101 cgggaaaaac tgaaggaagc atttaagcgt tttgatacga agaaaggtaa atgcctgacc
     2161 gataaagagg ctaaagaagc tggattctgt attaaaaaaa atgaacttgt gatgagcctg
     2221 aaatgcagca ttaagggtac tgggccggga cagatgatac gtattaataa caatgtgttc
     2281 aagactaatg tacataacgt gggcgtggac gtctatctgg atgaaaaagg caaaaaaaaa
     2341 gcttacgagc gtaaaaatcc ccggctgtca aaacacttta tagaacctcc gcctcagcca
     2401 aatgggcggg tgtccttcac cctgaaacgg cgtgatatgg tgaccgtaga aggggaagac
     2461 gctatatacc ggatcaagaa gcttggtacc tctcctacca tcgaggcagt ggttggctct
     2521 gatggcaaaa cacgcacggt ttcggcgaca aaattaacca aagcgaattc cgcagagtaa
     2581 taagcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     2641 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2701 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     2761 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2941 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3061 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3121 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3181 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     3241 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3301 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     3361 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     3421 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     3481 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     3541 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     3601 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     3661 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     3721 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     3781 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     3841 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     3901 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     3961 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     4021 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     4081 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
     4141 tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag
     4201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     5041 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     5101 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     5161 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5221 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5281 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     5341 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     5401 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     5461 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     5521 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     5581 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     5641 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     5701 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     5761 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     5821 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     5881 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
     5941 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     6001 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     6061 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     6121 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     6181 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     6241 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6301 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     6361 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     6421 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     6481 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     6541 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     6601 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     6661 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     6721 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     6781 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     6841 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     6901 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     6961 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     7021 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     7081 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     7141 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     7201 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     7261 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     7321 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     7381 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     7441 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     7501 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     7561 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     7621 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     7681 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     7741 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     7801 actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca cagcagcggc
     7861 ctggtgccgc gcggcagcca tatggctagc atgactggtg gacagcaaat gggtcgcgga
     7921 tccgaattc
//
