LOCUS       sequence-409760-        8186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dGeoCas9-mediated silencing of genomic gene in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8186)
  TITLE     van der Oost E. coli CRISPR plasmids
REFERENCE   2  (bases 1 to 8186)
  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"
COMMENT     Sequence Label: pdGCas9_scr
FEATURES             Location/Qualifiers
     source          1..8186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     390..409
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(476..578)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(970..987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1104..1649)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1184..1203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5042..5066
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5073..5078,5079..5095,5096..5101))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5264..5283)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5489..5566
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5567..6649
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5586..5605)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        6710..6744
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(join(8002..8186,1..475))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
       61 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
      121 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
      181 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
      241 ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
      301 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
      361 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
      421 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
      481 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
      541 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
      601 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
      661 cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga
      721 tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc
      781 tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc
      841 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
      901 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
      961 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     1021 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     1081 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     1141 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1201 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
     1261 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
     1321 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
     1381 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
     1441 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
     1501 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
     1561 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
     1621 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
     1681 tgtcgacagg attaaacaaa aatggcccgc ttcataagca ggccattttg tctgatttag
     1741 tcacgagtag attgcagtgg acgaatggtt ttgccaggtt tgctgtgtgc ggaggacgcc
     1801 agaccgacac gtttctcgcc acgcacttta tagatgttac ccagaacatc aacttgatat
     1861 ttctcgaaac gtttcagggt gcgagagcca acgccacgca gagagaaacg gtggtcgtga
     1921 gaaatcagtt ccaggccgcc gtttgcggag tcgatcgttt tatagtaaac gaacacgtct
     1981 ttcacgttaa tttcttcgcc tgcagcggtt ttaacggttt tttcacgcgg cagttcgatg
     2041 cggatcaggt cgttcggata caggctgaaa cgaaacgtgt aatcttcggt catttctttc
     2101 cattcagagt acggcttgtt cggttcaatc gctttgttcg gcagaatgcc tttcatgatg
     2161 tccatggtat acaccggtac acagtagtat ttaccatctt tttcgaacac gtccacgcga
     2221 acgatgttgg aattgtaggc cacggtttta ccatcgttca gagggatcac ctggtttttc
     2281 gtgtcgataa tttttacagt acggattacc gggcccggtt cgccgttttt tttcggtttg
     2341 tacagaggct cctggaatgc ctttttcggg tcgttgttat gctccagcag acgctggcgg
     2401 atggcttcat aagtgcgtgg atcggattct ttaccgtaca ttgggaagtg gccggatgca
     2461 tccagtttaa tctcggacag tttggttttt actacggtct ggattttacc ggagcgttcg
     2521 tcgatgccca cgtaacggcg cagcgtttcc tggtgcgcag cgccagtaac agaacgtttc
     2581 ggcatacggc tgacaaacac cggctgcaga gattccagtt tttggtcatc gtagttaccc
     2641 aggttcagag ctttgatgct ttctttagga tgtttggaca gacgagcacg cagttcgtcc
     2701 gcgaagtgcg gccacggctg cggaaaatgc ggttcggttt ttttggccag ttccttgttc
     2761 tgctcacgac gctggtagaa tgcggtcacc ttagcgatat cgcttggggt ggtgcacgcc
     2821 acaataacgg cgtcaacagc gtggtgcaga tcgctttcct cacggttctt gttgaactcc
     2881 cagcgagaac gcaggtgggc ggtaacacgg ccgtttacgg tataaacttt ctgtttatcg
     2941 tcggattccg caaatttcag gtgttcacga atgaagtttg cgaagaagcg gctgatgtag
     3001 cgagtatcgt tcaggttacg gtttttgaat tcagtctctt cgttttcatc gtagtgcaga
     3061 cgcagcagac ggtcacgttt ctttttagag aactgtttat tggtcaggac gaaggtttcg
     3121 aactgttgcc aacgctcagt gccaacaccc agatattcag ctgggatgcg gttgcctttc
     3181 tcacggtttt cgcgagtcag aaccagaact ttgttggtat aagaatcatc cagagagcgg
     3241 gagtacggga tcacagcatc tacttcaacg taacctggct ccagcagacg ttcgatctcg
     3301 atcggctgca gagagtaagc acagcgaccg ttctgttcag accacagctt gaacttcaca
     3361 atatcgtgac cagtagggtt cagagtcaga ccgtattcca tcagctggcg gatcgcggtt
     3421 tcgttttttt tacggttttc atcctgttcc tttttagttt tacgacgctc gtcaaaagtc
     3481 tggctcaggt cgcgcgccag ttcgatatgg atggaaaccg ggctgccgta ctttttgatg
     3541 atcgcgttga ctactttgcg cgcctgggtc agtgcacgca ttactacagg gttcgcaatc
     3601 ggcgggatgt tcggcagcag catagttttt tgcttctttt tcggaccggt gaaggtgtag
     3661 cctgcacgtt cacaagcgga ggagtagact tcaccctgtt ccatatacgg caggatggaa
     3721 cgcagagctt tcagagacag gtgaccgaat ttagtgaagg acaggttcag cagttcttca
     3781 atcagctcgt tatcgtaaac tttgttagcc aggttaggca tacgtttgcc gttctgttcg
     3841 tactcgttgc gcaggtaaga gtggatatcc gcatcgtctt taaacagggt cagcgcgtaa
     3901 ccgaaggtat caaagtcgat cggcaggaag ctgctggatt tacccttgcc gtaaacttta
     3961 tctacagctt tacggatctg gtggtatgca tccagttcca ggaaacgaat gttttcgttc
     4021 tgcttacgag attcaccgcg atcgtaaacg atgcccttga agtaggtgtc gtccggcaga
     4081 tgcagcaggg tacgaatatc gtggtaggta attttgtttt tctggaatgc ttgttcatac
     4141 agcagacgac gttcttcgtc ggtcaggccg cgggcgccag acggagaaat cagacgcagt
     4201 ttgttgatat gttcccacgc gataaaagac tggaaggtat aggtggcttt cggcgcacgt
     4261 ttttccttcg gttcgaaagt acaaaagcct actttttttt cgatatcatc tttagacgcc
     4321 accggacgct gagatgccca aatggtaata tattcgtttt cgaattcttc ggtgcaagac
     4381 atgttgccga attcgcgctg tttggagaaa atcagacgga tttcacgttc caggtcatcg
     4441 cgtgcaatgg tgttggtgta gttttcgcct ttgttacgct tatgcagtgc gaatttcggg
     4501 tctttaacaa tcatttcgcc cacggtacgg tagctagaca gaattgcacg gttctcctca
     4561 atatgtttca gcatagtgga gttctcttta ttggagcgtt cagatttacg gttagatttg
     4621 aagccacgac gctttgccag atgcagcagg acacgcgcca gttcgtcgtt gttcagctta
     4681 cggtccaggg cttctacacg cagctgccat acgtcgatct cgtgtttttc ttcgaacagt
     4741 ttatccagtt cttctttcgt caggatgcct tcacgaataa ccaggcgacg aatacgctcc
     4801 agacggtgtt tacgacgacg caggcggcgg cgtgcgctac gtgccagacg acgcggcaga
     4861 gccagagatt caccggtctg cgggttctcc gcacggtcaa aaatgcgcac gcccagatct
     4921 tcgatacgag ggatatccag gttcatgact gcccaaccaa cagaggtaat accgatggcc
     4981 aggccaatct tataacgcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa
     5041 cggaattgtt atccgctcac aattcctacc gcattaaagc ttatcgatga taagctgtca
     5101 aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     5161 taaattgcac tgaaatctag atttcgcggg atcgagatct cgatcctcta cgccggacgc
     5221 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     5281 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     5341 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     5401 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     5461 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
     5521 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     5581 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     5641 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     5701 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     5761 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     5821 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     5881 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     5941 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     6001 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     6061 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     6121 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     6181 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     6241 ttttcaacaa accatgcaaa tgctgaatga gggtatcgtt cccactgcga tgctggttgc
     6301 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     6361 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     6421 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     6481 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     6541 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6601 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgat cagaatcgac
     6661 tgatggacca aaaaaccccg ccgaagcggg gactttgaac tagcatacat tgacagctag
     6721 ctcagtccta ggtataatgc tagcctagat ccgcagtaac cccatgggtc atagttcccc
     6781 tgagattatc gctgtggtat aatgaaagtt ataccacagc aatgatctca gggttactat
     6841 gataagggct ttctgcctat aggcagactg acccgtggcg ttggggatcg cctatcgccc
     6901 gctttcttcg ggcattcccc actcttaggc gaacgaagac cttggcggaa aacgtcaagg
     6961 tcttttttat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
     7021 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     7081 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
     7141 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     7201 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     7261 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     7321 ggaaacgcgg aagtccccta cgtgctgctg aagttgcccg caacagagag tggaaccaac
     7381 cggtgatacc acgatactat gactgagagt caacgccatg agcggcctca tttcttattc
     7441 tgagttacaa cagtccgcac cgctgccggt agctccttcc ggtgggcgcg gggcatgact
     7501 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     7561 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgccc
     7621 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     7681 acatctgtat taacgaagcg ctaaccgttt ttatcaggct ctgggaggca gaataaatga
     7741 tcatatcgtc aattattacc tccacgggga gagcctgagc aaactggcct caggcatttg
     7801 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     7861 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
     7921 tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca
     7981 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     8041 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     8101 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     8161 gaagaagttg tccatattgg ccacgt
//
