LOCUS       V28C_M151E_ABEx4_SpRY   8217 bp    DNA     circular SYN 07-NOV-2025
DEFINITION  V28C_M151E_ABEx4_SpRY.
ACCESSION   V28C_M151E_ABEx4_SpRY
VERSION     V28C_M151E_ABEx4_SpRY
KEYWORDS    .
SOURCE      
  ORGANISM  .
            .
COMMENT     Annotated with pLannotate v1.2.2
FEATURES             Location/Qualifiers
     CDS             complement(1403..5504)
                     /note="pLannotate"
                     /label="dCas9 (fragment)"
                     /database="snapgene"
                     /identity="99.2"
                     /match_length="99.9"
                     /fragment="True"
                     /other="CDS"
                     /description="catalytically dead mutant of the Cas9
                     endonuclease from the Streptococcus pyogenes Type II
                     CRISPR/Cas system RNA-guided DNA-binding protein that lacks
                     endonuclease activity due to the D10A mutation in the RuvC
                     catalytic domain and the N863A mutation in the HNH
                     catalytic domain"
     enhancer        complement(6433..6736)
                     /note="pLannotate"
                     /label="CMV enhancer"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
                     /description="human cytomegalovirus immediate early
                     enhancer"
     CDS             7187..8047
                     /note="pLannotate"
                     /label="AmpR"
                     /database="snapgene"
                     /identity="99.8"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
                     /description="β-lactamase; bla; confers resistance to
                     ampicillin, carbenicillin, and related antibiotics"
     polyA_signal    complement(1062..1286)
                     /note="pLannotate"
                     /label="bGH poly(A) signal"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="polyA_signal"
                     /description="bovine growth hormone polyadenylation signal"
     promoter        complement(6229..6432)
                     /note="pLannotate"
                     /label="CMV promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /description="human cytomegalovirus (CMV) immediate early
                     promoter"
     rep_origin      1..589
                     /note="pLannotate"
                     /label="ori"
                     /database="snapgene"
                     /identity="99.8"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
                     /description="high-copy-number ColE1/pMB1/pBR322/pUC origin
                     of replication "
     promoter        7082..7186
                     /note="pLannotate"
                     /label="AmpR promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /description="bla"
     CDS             complement(5607..6098)
                     /note="pLannotate"
                     /label="TADA_ECOLI"
                     /database="swissprot"
                     /identity="87.8"
                     /match_length="98.2"
                     /fragment="False"
                     /other="CDS"
                     /description="Protein evidence at protein level: Swiss-Prot
                     protein existence level 1. Catalyzes the deamination of
                     adenosine to inosine at the wobble position 34 of
                     tRNA(Arg2). Essential for cell viability. From Escherichia
                     coli (strain K12)."
     promoter        913..943
                     /note="pLannotate"
                     /label="lac promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /description="promoter for the E. coli lac operon"
     protein_bind    877..898
                     /note="pLannotate"
                     /label="CAP binding site"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="protein_bind"
                     /description="CAP binding activates transcription in the
                     presence of cAMP. E. coli catabolite activator protein"
     promoter        complement(6169..6187)
                     /note="pLannotate"
                     /label="T7 promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /description="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    951..967
                     /note="pLannotate"
                     /label="lac operator"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="protein_bind"
                     /description="The lac repressor binds to the lac operator
                     to inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG). lac repressor
                     encoded by lacI"
     CDS             772..864
                     /note="pLannotate"
                     /label="lacI (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="8.6"
                     /fragment="True"
                     /other="CDS"
                     /description="lac repressor; lacI; The lac repressor binds
                     to the lac operator to inhibit transcription in E. coli.
                     This inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
     repeat_region   complement(749..760)
                     /note="pLannotate"
                     /label="AAV2 ITR (fragment)"
                     /database="itrs"
                     /identity="100.0"
                     /match_length="8.3"
                     /fragment="True"
                     /other="repeat_region"
                     /description="AAV2 ITR Flip"
     misc_feature    complement(1016..1032)
                     /note="pLannotate"
                     /label="MCS (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="15.7"
                     /fragment="True"
                     /other="misc_feature"
                     /description="pBluescript multiple cloning site"
     promoter        complement(1037..1058)
                     /note="pLannotate"
                     /label="SV40 promoter (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="6.1"
                     /fragment="True"
                     /other="promoter"
                     /description="SV40 enhancer and early promoter"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccctag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtatc gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
     1081 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     1141 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     1201 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     1261 agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttaaa ctcaatggtg
     1321 atggtgatga tgaccggtta gactttcctc ttcttcttgg gctcgaattc gctgccgtcg
     1381 gcggttcttt ttgagccgcc agagtcacct cccagctgag acaggtcgat ccgtgtctcg
     1441 tacaggccgg tgatgctctg gtggatcagg gtggcgtcca gcacctcttt ggtgcttctg
     1501 tactgcttgg ggtcgatggt ggtgtcaaag tacttgaagg ctctaggggc tcccagtctg
     1561 gtcagggtaa acaggtggat gatattctcg gcctgctctc tgatgggctt atcccggtgc
     1621 ttgttgtagg cggacagcac tttgtccaga ttagcgtcgg ccaggatcac tctcttggag
     1681 aactcgctga tctgctcgat gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc
     1741 tgtttctgct cattatcctc gggggagccc ttcagcttct catagtggct ggccaggtac
     1801 aggaagttca catatttgga gggcagggcc agttcgtttc ccttctgcag ctgcttggca
     1861 gaggccagca ttctcttccg gccgttttcc agctcgaaca gggagtactt aggcagcttg
     1921 atgatcaggt cctttttcac ttctttgtag cccttggctt ccagaaagtc gatgggattc
     1981 ttctcgaagc tgcttctttc catgatggtg atccccagca gctctttcac actcttcagt
     2041 ttcttggact tgcccttttc cactttggcc accaccagca cagaataggc cacggtgggc
     2101 cacaggaagc cgccgtactt cttagggtcc cagtccttct ttctggcgat cagcttatcg
     2161 ctgttcctct tgggtctgat agactctttg ctgaagccgc ctgtctgcac ctcggtcttt
     2221 ttcacgatat tcacttgggg catgctcagc actttccgca cggtggcaaa atcccggccc
     2281 ttatcccaca cgatctcccc ggtttcgccg tttgtctcga tcagaggccg cttccggatc
     2341 tcgccgttgg ccagggtaat ctcggtcttg aaaaagttca tgatgttgct gtagaagaag
     2401 tacttggcgg tagccttgcc gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg
     2461 tacaccttgt agtcgccgta cacgaactcg ctttccagct tagggtactt tttgatcagg
     2521 gcggttccca cgacggcgtt caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg
     2581 cgcactttgt aaaactggaa atccttccgg aaatcggaca ccagcttgga cttcagggtg
     2641 atcactttca cttcccggat cagcttgtca ttctcgtcgt acttagtgtt catccgggag
     2701 tccaggatct gtgccacgtg ctttgtgatc tgccgggttt ccaccagctg tctcttgatg
     2761 aagccggcct tatccagttc gctcaggccg cctctctcgg ccttggtcag attgtcgaac
     2821 tttctctggg taatcagctt ggcgttcagc agctgccgcc agtagttctt catcttcttc
     2881 acgacctctt cggagggcac gttgtcgctc ttgccccggt tcttgtcgct tctggtcagc
     2941 accttgttgt cgatggagtc gtccttcaga aagctctgag gcacgatatg gtccacatcg
     3001 tagtcggaca gccggttgat gtccagttcc tggtccacgt acatatcccg cccattctgc
     3061 aggtagtaca ggtacagctt ctcgttctgc agctgggtgt tttccacggg gtgttctttc
     3121 aggatctggc tgcccagctc tttgatgccc tcttcgatcc gcttcattct ctcgcggctg
     3181 ttcttctgtc ccttctgggt ggtctggttc tctctggcca tttcgatcac gatgttctcg
     3241 ggcttgtgcc ggcccatcac tttcacgagc tcgtccacca ccttcactgt ctgcaggatg
     3301 cccttcttaa tggcggggct gccggccaga ttggcaatgt gctcgtgcag gctatcgccc
     3361 tggccggaca cctgggcttt ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc
     3421 agctgcatga agtttctgtt ggcgaagccg tcggacttca ggaaatccag gattgtcttg
     3481 ccggactgct tgtcccggat gccgttgatc agcttccggc tcagcctgcc ccagccggtg
     3541 tatctccgcc gcttcagctg cttcatcact ttgtcgtcga acaggtgggc ataggttttc
     3601 agccgttcct cgatcatctc tctgtcctca aacagtgtca gggtcagcac gatatcttcc
     3661 agaatgtcct cgttttcctc attgtccagg aagtccttgt ccttgataat tttcagcaga
     3721 tcgtggtatg tgcccaggga ggcgttgaac cgatcttcca cgccggagat ttccacggag
     3781 tcgaagcact cgattttctt gaagtagtcc tctttcagct gcttcacggt cactttccgg
     3841 ttggtcttga acagcaggtc cacgatggcc tttttctgct cgccgctcag gaaggcgggc
     3901 tttctcattc cctcggtcac gtatttcact ttggtcagct cgttatacac ggtgaagtac
     3961 tcgtacagca ggctgtgctt gggcagcacc ttctcgttgg gcaggttctt atcgaagttg
     4021 gtcatccgct cgatgaagct ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc
     4081 cagggggtga tggtttcctc gctctttctg gtcatccagg cgaatctgct gtttcccctg
     4141 gccagagggc ccacgtagta ggggatgcgg aaggtcagga tcttctcgat cttttcccgg
     4201 ttgtccttca ggaatgggta aaaatcttcc tgccgccgca gaatggcgtg cagctctccc
     4261 aggtggatct ggtgggggat gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc
     4321 tctctgttca gcttcacgag cagttcctcg gtgccgtcca tcttttccag gatgggcttg
     4381 atgaacttgt agaactcttc ctggctggct ccgccgtcaa tgtagccggc gtagccgttc
     4441 ttgctctggt cgaagaaaat ctctttgtac ttctcaggca gctgctgccg cacgagagct
     4501 ttcagcaggg tcaggtcctg gtggtgctcg tcgtatctct tgatcataga ggcgctcagg
     4561 ggggccttgg tgatctcggt gttcactctc aggatgtcgc tcagcaggat ggcgtcggac
     4621 aggttcttgg cggccagaaa caggtcggcg tactggtcgc cgatctgggc cagcaggttg
     4681 tccaggtcgt cgtcgtaggt gtccttgctc agctgcagtt tggcatcctc ggccaggtcg
     4741 aagttgctct tgaagttggg ggtcaggccc aggctcaggg caatcaggtt tccgaacagg
     4801 ccattcttct tctcgccggg cagctgggcg atcagatttt ccagccgtct gctcttgctc
     4861 agtctggcag acaggatggc cttggcgtcc acgccgctgg cgttgatggg gttttcctcg
     4921 aacagctggt tgtaggtctg caccagctgg atgaacagct tgtccacgtc gctgttgtcg
     4981 gggttcaggt cgccctcgat caggaagtgg ccccggaact tgatcatgtg ggccagggcc
     5041 agatagatca gccgcaggtc ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg
     5101 tagatggtgg ggtacttctc gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg
     5161 tgccgctcgt gcttcttatc ctcttccacc aggaaggact cttccagtct gtggaagaag
     5221 ctgtcgtcca ccttggccat ctcgttgctg aagatctctt gcagatagca gatccggttc
     5281 ttccgtctgg tgtatcttct tctggcggtt ctcttcagcc gggttctctc ggctgtttcg
     5341 ccgctgtcga acagcagggc tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg
     5401 cccagcacct tgaatttctt gctgggcacc ttgtactcgt cggtgatcac ggcccagccc
     5461 acagagttgg tgccgatggc caggccgatg ctgtacttct tgtctgaccc cccgctgctg
     5521 cccccgctgc tttcaggtgt tgcgctctcg cttgtgccag gtgtctcaga gccagaggag
     5581 cctccgctag atcctccgga gttgatggag ctctgggcct tcttctgagc attgaacacc
     5641 tgtctaggtt cccgatagaa atcgcacagc agggcggcac attcatctgc caggattccc
     5701 tcggtaattt cgacgcggtg attcatgccg gggtagttca gcacgttcat cagggagcct
     5761 gcggcgcctc tttttgagtt cctcacgcca aacaccacgc ggccgatcct agagtggatc
     5821 atggcgccgg cgcacatcac gcaaggctcg aatgtcacgt acagggtggc gtcaatcagt
     5881 ctgtagttct gcatgaccag gccgccctgt ctcagggcca taatttcggc atgggctgtt
     5941 gggtcgtgca ggccgatggc tctgttccag ccctcgccga tcactctatt gttcagcacc
     6001 agcacggctc ccacagggca ctccctctca tcccgtgccc tcttggccag ggtcagggca
     6061 tgtctcatcc agtactcgtg ggaaaactcc acctcagaga ctttccgctt cttctttggt
     6121 gactcgaact cgcttccgtc ggctgtccgt ttcatggtgg cggctctccc tatagtgagt
     6181 cgtattagcg gccgcggatc tctagcggat ctgacggttc actaaaccag ctctgcttat
     6241 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     6301 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     6361 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     6421 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     6481 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     6541 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     6601 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     6661 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     6721 cgtaagttat gtaacgcgga actcaatata tgggctatga actaatgacc ccgtaattga
     6781 ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg
     6841 aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg
     6901 ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg
     6961 gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag tcgaccctag
     7021 gggatcggga gatcgatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     7081 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7141 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7201 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7261 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7321 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7381 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7441 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7501 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7561 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7621 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7681 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7741 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7801 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7861 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7921 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7981 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8041 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8101 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8161 acgtgagttt tcgttccact gagtgtcaga ccccgtagaa aagatcaaag gatcttc
//
