LOCUS       ancCas9d_pET28a         7932 bp    DNA     circular UNA 13-MAY-2025
DEFINITION  Generated by Golden Gate/Type IIS Cloning (34-1 in pET-28a(+) PCR
            Product, MG34-29_in_pET-21(+) PCR Product).
ACCESSION   urn.local...2s4-j5vvyr0
VERSION     urn.local...2s4-j5vvyr0
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          join(1..>336,<2581..7932)
                     /organism="34-1 in pET-28a(+)"
     misc_feature    333..2584
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="MG34-29_in_pET-21(+) PCR Product
                     (concatenated sequence 2)"
     source          <337..>2580
                     /organism="MG34-29 in pET-21(+)"
     misc_feature    <337..2580
                     /note="Geneious type: misc_feature_"
                     /standard_name="insert"
     misc_feature    2581..336
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="34-1 in pET-28a(+) PCR Product
                     (concatenated sequence 1)"
     misc_feature    2787..3093
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: rep_origin"
                     /standard_name="f1"
     misc_feature    3317..4132
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: Antibiotic R"
                     /standard_name="kan"
     primer_bind     complement(3688..3713)
                     /standard_name="KanR_Rv_Primer_(3)"
     primer_bind     3714..3744
                     /standard_name="KanR_Fw_Primer_(1)"
     misc_feature    4254..4842
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: rep_origin"
                     /standard_name="colE1_high_copy"
     misc_feature    5269..5460
                     /note="Geneious type: misc_feature_"
                     /standard_name="ROP"
     misc_feature    6235..6256
                     /note="Geneious type: misc_feature_"
                     /standard_name="CAP_binding"
     misc_feature    6269..7360
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: ROI"
                     /standard_name="lacI"
     misc_feature    7352..7429
                     /note="Geneious type: misc_feature_"
                     /standard_name="lacIQ_promoter"
     misc_feature    7738..7756
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: promoter"
                     /standard_name="T7"
     misc_feature    7756..7783
                     /note="Geneious type: misc_feature_"
                     /standard_name="lacO"
     misc_feature    7796..7818
                     /note="Geneious type: misc_feature_"
                     /note="Geneious type: RBS"
                     /standard_name="RBS_3"
     misc_feature    7837..7853
                     /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 agaaggagct ggtgttaggt
      361 attgactatg gcgggaagta cactggtctg gcagtagtgg atagacgcaa caatcaagta
      421 ctgtatgccc ggacggttaa gatgcgagac gacgttgccg acatacttaa aggtcgtcgc
      481 gagcaaagag gtattagacg tacccaacag acaaagaaga agcgtctccg cgagttgaag
      541 aagtacttaa actcgatcgg actgtatgac gagtccacgg aaacatttaa gaccatctac
      601 agtctcgcgc acaaaagagg ctacgactat gcggacatgc cagaggagaa gaccagcgaa
      661 gagatagaag agatggacga ggaagaacgc aagcaatggg aaaaggagaa gaaagagtgg
      721 gaagagacgc aacgtaacag tagacatcgt gaagaggtcc tgaaggatgt tcgaaaagtt
      781 atgagtgagg ggaacgcaag tgaggagcaa attaaacggg tagagtccat cttcaacaaa
      841 cagtatcgcc ctaagcgttt caataaccgt atcctcacga agtgcaaagt ttgtgggaag
      901 aacacccctc tccgcagaaa tgttcgagag ttgttactgg aaaatatagt gagattctta
      961 cctctggagt ctgaagagaa ggagacgctg aagcaaacca tactggagaa aggccaacag
     1021 gagaatatca acaagatatt tcgcaagttg aagttcaacc agaaggattg gccaggaaag
     1081 aaccttacag acattgcaaa gaataagtta cctggacgtc tccctttctg taaagagcat
     1141 ttcgcagaga acgagaagtt cacaactata gagaaatcta ctttccgcct ggcaccgtcg
     1201 ctgaagacta agattgaaaa tgtgcttaca gtgataaagg acgaaatcct gccaaatttc
     1261 actcttaaca aagtggtgat ggaaagcaac aacttcgaca tcgcagccaa aacgaagggc
     1321 aagaagcgtc ttgcaaagga agagtattca aagggacacc gcgagaacaa ggagacttta
     1381 aaggaagcat tactgagaga aacggacggc cgctgtgtgt actgtggaaa atcaattgac
     1441 ctgtctaacg cgaacaaaga ccacatatac cctagaaagg ccggcggtct taacattttc
     1501 gctaatctcg ttgcatgctg ccgcagttgt aatgagaaca agaaggggcg tacaccgctt
     1561 gaaagcggga taatgcccaa gcctgaagtg gtagccacta tcaagaatga cctgaagaag
     1621 aagatattgg aagatgcgca acaaattaac accctggact tcaataagta tatgagtcat
     1681 gcgtcgattg gttggcgcca tatgcgcgat cggcttcgag aactgaccgg aaataagaag
     1741 ctgccaatcg agcgccaatc aggaatagta actgcatact tccgccgctg gtggggtttc
     1801 aagaaagagc gtgggaatga caaacaccat gcgttggacg cagttattct tgcctcgcgt
     1861 aaggactact cagacgacgg ccttgttgat atgacactta agcctgcaaa caacgacggc
     1921 cgtgagttcg acccagagaa gcacatcaaa gagatgaagg agttcaagcg cgacaagggt
     1981 tccaagggat ccgcgctgca cgatcgaaac cccttatcat acaagaatgg aataatcacg
     2041 cggcgcttca tggtaactga gattgagcgc ggcaaggaag acgcggtcat atcagaagag
     2101 taccgtgaaa agcttaagga agcattcgat cggttcggta taaagaatgg gaagtgtctt
     2161 acggacaagc aggcgaaaga agcaggcctt tacatcaaga agaacgaggg tgttatgagc
     2221 cttaagtgca gtgttaaagg caccggtcca ggacaaatga taagaatcaa gaataacgtt
     2281 ttcaaaacta atgtacacaa tgtcggagta gcagtatact tggacgaaaa gggtaagaag
     2341 cgtgcatgtg aacttaagaa tccacgtctt tccaagcatt tcgtggaacc tcagcctcag
     2401 gtcaaaggca aaatcctctt caccttacgt cgtggcgaca tggtaactgt ggaaggtgga
     2461 gatgccatat acagaatcaa gaagttgggc acatccccag ttatcgagat cgttccaggg
     2521 tcggacggaa agactaagac agtatctgca acaaaattga ccaaggctaa caacgcgaag
     2581 taataagcgg ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     2641 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2701 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     2761 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2821 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2881 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2941 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3001 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3061 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3121 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3181 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3241 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3301 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     3361 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     3421 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     3481 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     3541 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     3601 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     3661 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     3721 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     3781 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     3841 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     3901 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     3961 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     4021 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     4081 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     4141 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     4201 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4381 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4921 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4981 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     5041 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     5101 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     5161 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5221 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5281 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     5341 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5401 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5461 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     5521 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     5581 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     5641 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     5701 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     5761 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     5821 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     5881 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     5941 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     6001 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     6061 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     6121 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     6181 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     6241 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6301 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6361 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     6421 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     6481 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     6541 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     6601 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     6661 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     6721 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     6781 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     6841 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     6901 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     6961 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     7021 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     7081 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     7141 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     7201 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     7261 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     7321 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     7381 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     7441 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     7501 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     7561 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     7621 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     7681 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     7741 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     7801 ttaactttaa gaaggagata taccatgggc agcagccatc atcatcatca tcacagcagc
     7861 ggcctggtgc cgcgcggcag ccatatggct agcatgactg gtggacagca aatgggtcgc
     7921 ggatccgaat tc
//
