LOCUS       sequence-316111-        8739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPRi. Arabinose-inducible ddcpf1 gene with CRISPR array 
            lacking a gene-specific spacer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8739)
  AUTHORS   Jervis AJ, Hanko EKR, Dunstan MS, Robinson CJ, Takano E, Scrutton NS
  TITLE     A plasmid toolset for CRISPR-mediated genome editing and CRISPRi 
            gene regulation in Escherichia coli.
  JOURNAL   Microb Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780.
  PUBMED    33710766
REFERENCE   2  (bases 1 to 8739)
  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"; journalName: "Microb 
            Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780."
COMMENT     Sequence Label: pBbS8c-ddcpf1-Delta
FEATURES             Location/Qualifiers
     source          1..8739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2056..2340)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(2114..2133)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2367..3245
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3275..3377
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3378..4037
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3444..3463)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4058..4152
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4914..5864
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(6395..6422)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6438..6509)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    6597..6613
                     /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(6621..6627,6628..6644,6645..6650))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      7030..7039
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttgttcaggt cgtaaatttc tttggttatt tccagcggtt ctataaagtt atttgaaagc
       61 aggatcgggg tggtgtgtgt ctgaaaatgc gcggtcaccg ctttcagctg ggtcgaacac
      121 ttcggaatca ttttggcggc atccggaaag tagtcgtaat acattttatc aaacccttca
      181 ctggtcttct ccgtcggttc gaatgacagt gccttgtaac ggcctttctg cttcggcata
      241 atgcccagat aatacaggcc gtttttcaca aataagatcg caccgttgtt cttctctttg
      301 ttgacatccc aacctgacgc tagggtcggc atttggaagt tcaatttgaa tttttccact
      361 gaataaggct ttttggtcgc atagttgcgt gctttgttgt aaaagctcaa cgacggttcc
      421 atctccaatt ttataccggt caggcgtgcg ctaaattcag ggtccacctc attagactcg
      481 tccaccgcaa accagtccag caagtggtac agtcccagta gagaatctaa ttgagatttc
      541 aggatttctt tctcttcctg ctttttcagc gtcgtcggca gaggctgatc tagggccgcg
      601 tgcgcatgtg agaggatttc gcttgtcttc tgtttgaaag cctccgacag ttccttaccg
      661 gccgccgaaa taatctcttg taagttaata tcttcatgtt tgagggagcg ctggactttc
      721 tctttagctg acttggtgat ctttccggtc agctcggaaa ttcgacgttc atacagggca
      781 tttctcagcg tgtcccaatg gtcgcacagc gcgctgctga tagtttccag ctttttatgt
      841 gaaataaaaa tatgggtgag atcaatgctg ttcagttcgt taaagagggc ctcagccgtt
      901 tccaggacgt tttcattgcg cagtaacgtt ttatatttgc agaagctctg aattacttct
      961 tcatcgcttt taaattcctc caggataaag ctcagtgtgt tgcggtcgga cagaatttgt
     1021 ttaaagagcg gtataaatcg atgggggaga gatgcaataa tgtgcgctgt ttcatcgttc
     1081 ttttgaattg ccaggttcag cacctcattc aggcctttta tcttttccgt tccagcttct
     1141 cggctaatac cacctaagag ctgattatat aggtcaatct gtgtctgggt aagaagctga
     1201 ttatagaacg ggaatgaaaa cacttcttca atgctggttg acacgaaaat gccaatcgcc
     1261 ttcttaacat tttcaaaatg ttctcgcagt gatggcactg ctgtgatcag acgagtaaaa
     1321 atatgacaat tttctttaaa tttcggaaaa ttgtcttgaa caatacgatg cggaatggcc
     1381 gtactaatat cttctgcgct aaacacgttc ttgcgattct cgtagaaccc ggaaaaatac
     1441 gtagtgaatt tatcaaaaga acgcagcaag gcattttcat gttccgtggt cgttacggta
     1501 cctaactgct ttaatacttt accgttgaac agctccgctt taaacagacc tttgtatatt
     1561 tcggcatgac gcttgttgat ggcgtccgtt agattatctg tcctaccaat aaaatagtca
     1621 tgaattgcat tgcggtatgt agcctgctcc tcgattaatg cattacgagt ttcttctgtt
     1681 ttctccttac ggtaagagtc tatagccgcg cttaaatttt cccaatctaa ctgcaccagt
     1741 tgcagacact ggtctgcata cgtcttgtag atgcgatcga taatcggttt aagttcttta
     1801 taatggtcgt ttcgggcctt atcctcttca atgaagcctt gctcttgaat gtgtttcagt
     1861 gttttgccct ggggaatcag ctcaaatctc agcgtcttgc ttacctgata aagattggta
     1921 aaaccttcaa actgggtcat cacgacctcc tacataaaaa cagtttcatg gattatgaac
     1981 cgcgcaagag aataacactg acatgcgagg cattttatga cggtgtagat cttttgaatt
     2041 cccaaaaaaa cgggtatgga gaaacagtag agagttgcga taaaaagcgt caggtagaat
     2101 ccgctaatct tatggataaa aatgctatgg catagcaaag tgtgacgccg tgcaaataat
     2161 caatgtggac ttttctgccg tgattataga cacttttgtt acgcgttttt gtcatggctt
     2221 tggtcccgct ttgttacaga atgcttttaa taagcggggt taccggtttg gttagcgaga
     2281 agagccagta aaagacgcag tgacggcaat gtctgatgca atatggacaa ttggtttctt
     2341 ctctgaatgg cgggagtatg aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg
     2401 gatactcgtt taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc
     2461 tcgatttttt tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc
     2521 gcggtcaggg ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt
     2581 tgctgttccc gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt
     2641 atcaccagtg ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt
     2701 caatatttgc caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg
     2761 acctgtttgg gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg
     2821 cgataaatct gcttgagcaa ttgttactgc ggcgcatgga agcgattaac gagtcgctcc
     2881 atccaccgat ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag
     2941 acagcaattt tgatatcgcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt
     3001 cacatctttt ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta
     3061 tcagccaggc gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca
     3121 atgttggttt tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca
     3181 gcccgagcga gttccgtgcc ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt
     3241 cataagacgt cgatatctgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     3301 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     3361 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     3421 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     3481 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     3541 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta
     3601 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     3661 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     3721 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     3781 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     3841 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     3901 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     3961 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     4021 ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta
     4081 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     4141 attggtgaga atccaagcac tagggacagt aagacgggta agcctgttga tgataccgct
     4201 gccttactgg gtgcattagc cagtctgaat gacctgtcac gggataatcc gaagtggtca
     4261 gactggaaaa tcagagggca ggaactgctg aacagcaaaa agtcagatag caccacatag
     4321 cagacccgcc ataaaacgcc ctgagaagcc cgtgacgggc ttttcttgta ttatgggtag
     4381 tttccttgca tgaatccata aaaggcgcct gtagtgccat ttacccccat tcactgccag
     4441 agccgtgagc gcagcgaact gaatgtcacg aaaaagacag cgactcaggt gcctgatggt
     4501 cggagacaaa aggaatattc agcgatttgc ccgagcttgc gagggtgcta cttaagcctt
     4561 tagggtttta aggtctgttt tgtagaggag caaacagcgt ttgcgacatc cttttgtaat
     4621 actgcggaac tgactaaagt agtgagttat acacagggct gggatctatt ctttttatct
     4681 ttttttattc tttctttatt ctataaatta taaccacttg aatataaaca aaaaaaacac
     4741 acaaaggtct agcggaattt acagagggtc tagcagaatt tacaagtttt ccagcaaagg
     4801 tctagcagaa tttacagata cccagatcac ccgggaaaag gactagtaat tatcattgac
     4861 tagcccatct caattggtat agtgattaaa atcacctaga ccaattgaga tgtatgtctg
     4921 aattagttgt tttcaaagca aatgaactag cgattagtcg ctatgactta acggagcatg
     4981 aaaccaagct aattttatgc tgtgtggcac tactcaaccc cacgattgaa aaccctacaa
     5041 ggaaagaacg gacggtatcg ttcacttata accaatacgc tcagatgatg aacatcagta
     5101 gggaaaatgc ttatggtgta ttagctaaag caaccagaga gctgatgacg agaactgtgg
     5161 aaatcaggaa tcctttggtt aaaggctttg agattttcca gtggacaaac tatgccaagt
     5221 tctcaagcga aaaattagaa ttagttttta gtgaagagat attgccttat cttttccagt
     5281 taaaaaaatt cataaaatat aatctggaac atgttaagtc ttttgaaaac aaatactcta
     5341 tgaggattta tgagtggtta ttaaaagaac taacacaaaa gaaaactcac aaggcaaata
     5401 tagagattag ccttgatgaa tttaagttca tgttaatgct tgaaaataac taccatgagt
     5461 ttaaaaggct taaccaatgg gttttgaaac caataagtaa agatttaaac acttacagca
     5521 atatgaaatt ggtggttgat aagcgaggcc gcccgactga tacgttgatt ttccaagttg
     5581 aactagatag acaaatggat ctcgtaaccg aacttgagaa caaccagata aaaatgaatg
     5641 gtgacaaaat accaacaacc attacatcag attcctacct acataacgga ctaagaaaaa
     5701 cactacacga tgctttaact gcaaaaattc agctcaccag ttttgaggca aaatttttga
     5761 gtgacatgca aagtaagtat gatctcaatg gttcgttctc atggctcacg caaaaacaac
     5821 gaaccacact agagaacata ctggctaaat acggaaggat ctgaggttct tatggctctt
     5881 gtatctatca gtgaagcatc aagactaaca aacaaaagta gaacaactgt tcaccgttac
     5941 atatcaaagg gaaaactgtc catatgcaca gatgaaaacg gtgtaaaaaa gatagataca
     6001 tcagagcttt tacgagtttt tggtgcattc aaagctgttc accatgaaca gatcgacaat
     6061 gtaacagatg aacagcatgt aacacctaat agaacaggtg aaaccagtaa aacaaagcaa
     6121 ctagaacatg aaattgaaca cctgagacaa cttgttacag ctcaacagtc acacatagac
     6181 agcctgaaac aggcgatgct gcttatcgaa tcaaagctgc cgacaacacg ggagccagtg
     6241 acgcctcccg tggggaaaaa atcatggcaa ttctggaaga aatagcgctt tcagccggca
     6301 aaccggctga agccggatct gcgattctga taacaaacta gcaacaccag aacagcccgt
     6361 ttgcgggcag caaaacccgt accctaggta taaacgcaga aaggcccacc cgaaggtgag
     6421 ccagtgtgac tctagtagag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     6481 tctttcgact gagcctttcg ttttatttga tgcctggaga tccttactcg agtttggatc
     6541 cgagtttttg ttcgggccca agcttcaaaa aaaatctaca agagtagaaa ttgaaattgt
     6601 tatccgctca caattccaca cattatacga gccggatgat taattgtcaa cagctcattt
     6661 aggtgtagac atcaagttac caaagttgcg cagcgaaaaa accccgccga agcggggttt
     6721 tttgcggtcc atgagggtga aggtcacatg gactatcaat tgcgcagttc ctgaatatac
     6781 gcgagccaat cctgatttga aatgccattc tgtagtttga ggtctttgga ttctttgaga
     6841 tgattcagta acagctggcc tttcagcgca atatggtatg ctccgttggc gtccgcgtcc
     6901 atcggccatt cggggttctg gaagcgacta tcaaaacaga cgccgttgag atcgcgtacc
     6961 gggctgttaa tataatcttc ccccgtcgcc gcgttagagt tacgcatttg aagaacgctg
     7021 cggataagag ccaccatggt atcaatcgcg tgcgagtcat cattttccaa cagtttaggc
     7081 agaatattgg aaccgtcacg aaatacaatt cccttttctt cgagcagtgc aatcagctca
     7141 ttagccgggt acaggtcgcg ataacggcct gtaaaccgat gattttcgat caccggtact
     7201 atacgtttcc cggcaataaa cggtgtgcct ttggcgtcga actgcgtttc attcttttcg
     7261 aacacaatat cccacgccgg cataaaccca ggcagaccgc gctgaaagct aagattgcga
     7321 ttcattttga agtgcagaat aaaatcgcct gttttcacat cgtagtgaag aaaatcaaag
     7381 ccctccagga aatgtttgcg gctttcgtgg tttttgatgg ttttccatac aaaaggatca
     7441 acaaagccgg tcaggggatc gattttgctg gtatacggtg cgggaacata aaacagaaaa
     7501 ccgctctgtg tacccatctt cgcaaagctg gtaaattgat ctgtaagctg gtacgggttg
     7561 agaacgccac ctaccttttc agcgggataa tcttttagta ctaagcaatt gagtttatcg
     7621 atcagcatct tctcgaactg ctgatacacc gccttttcgg caatgcccgt ccgctttgac
     7681 ttaaatccga agttcaggtt ggccagcacc acaacggctt gataatgtat cattaagtct
     7741 acgatttcgt gtataacctg ggataaataa ccttgtttaa gatccttaat cgtcccaact
     7801 accgaccaag cctgccgtgc cgccacacgt tctttttccc gattatctaa ctttttctgg
     7861 taatcaaatt gctgaatggt gtttaggcta cgctgttcta aaatcttgcc ggtagagtcg
     7921 attacggtta tatagataag attgcgttcg ccacggtcaa tcccaataat aggggtttcc
     7981 gggtgctctt tcaggtacgc gtttacgcgc tggttaaatt tcgagggtga gttcgccgcc
     8041 tggtaattaa gggtaatcgg tacgtggaaa aagaatttgt cggaggtaaa acgtcgatcc
     8101 ttaatgatct cgtgtgacac ctctttggtt attacgttcg gcagtaaggc ccgtgcttcg
     8161 tccgacagat cgtgactcag acggtgattg acatagtcgt aaagttcctg atacagcgtg
     8221 tcagggatcg gtgttttctg atccttcagt ttcttattga gcattttctc cccaagacga
     8281 tgggccatcc gcttcatgcg cgatttcgga cggtagaaca gctcggcttg gccattgagt
     8341 ttaattgagg tcttggccaa gttttcggga gagaacaggc cagtccaata gagagtatgt
     8401 agattcggtt tgccgtgatg gcctttcgca aaatccttgt tgtaaatctg gaacagataa
     8461 agctttccag tttcgactgc atccatgatt tctttttccg caatgcgctg gaagctaatg
     8521 tggtacagta acgggttcag ctcggcataa tattcgccca ggtctttata ctgggagctc
     8581 ggacgcagcg aactcaaatc tattgaagtg gtcttggtgt atttgctgag aaaatcgcgc
     8641 gtaaaatcaa tccatttaca cagggcttcg cggtaacctt tttgatctcc ggttttcttt
     8701 gcgtaggcgg tctgaaattt cttcggttct ttttccgga
//
