LOCUS       sequence-114422-        4676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial inducible gRNA expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4676)
  TITLE     Bacterial constitutive gRNA expression plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4676)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFG018
FEATURES             Location/Qualifiers
     source          1..4676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(11..889)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        916..1200
                     /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     1123..1142
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        1221..1296
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(1482..2432)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2480..2702
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        3257..3311
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3292..3310
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3319..4524
                     /codon_start=1
                     /gene="tetA from transposon Tn10"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGV
                     LLALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLS
                     GITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFI
                     AALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQL
                     IGQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLE
                     FIADSSAFAFLAFISEGWLDFPVLILLAGGGIALPALQGVMSIQTKSHEQGALQGLLVS
                     LTNATGVIGPLLFTVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQET
                     SA"
ORIGIN
        1 catcgattta ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
       61 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaag tagagttgat
      121 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
      181 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
      241 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
      301 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
      361 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
      421 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
      481 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
      541 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
      601 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
      661 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
      721 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
      781 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
      841 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
      901 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
      961 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     1021 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     1081 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     1141 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     1201 tcgcaagatg tggcgtgtta gttttagagc tagaaatagc aagttaaaat aaggctagtc
     1261 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt aaaaaaaaaa ccccctgtca
     1321 ggggcggggt ttttttttac aagtctcagt gtgctgaagt tttgggacca ttctccatgg
     1381 gtatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta
     1441 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     1501 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     1561 agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     1621 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat
     1681 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     1741 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     1801 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     1861 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     1921 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     1981 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     2041 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     2101 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     2161 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     2221 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     2281 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     2341 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     2401 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     2461 tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt cctttgagtt
     2521 gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     2581 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     2641 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     2701 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     2761 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcggttgcg
     2821 gccgcaatcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc
     2881 gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta
     2941 aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa
     3001 agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac
     3061 tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc
     3121 cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacgggg
     3181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag atatgtaacg
     3241 cacgttgtgc atcgatttga caccctatca ttgatagagt tattttccca ctccctatca
     3301 gtgatagaga aaagtgaaat gaatagttcg acaaagatcg cattggtaat tacgttgctc
     3361 gatgccatgg ggattggcct tatcatgcca gtcttgccaa cgttattacg tgaatttatt
     3421 gcttcggaag atatcgctaa ccactttggc gtattgcttg cactttatgc gttaatgcag
     3481 gttatctttg ctccttggct tggaaaaatg tctgaccgat ttggtcggcg cccagtgctg
     3541 ttgttgtcat taataggcgc atcgctggat tacttattgc tggctttttc aagtgcgctt
     3601 tggatgctgt atttaggccg tttgctttca gggatcacag gagctactgg ggctgtcgcg
     3661 gcatcggtca ttgccgatac cacctcagct tctcaacgcg tgaagtggtt cggttggtta
     3721 ggggcaagtt ttgggcttgg tttaatagcg gggcctatta ttggtggttt tgcaggagag
     3781 atttcaccgc atagtccctt ttttatcgct gcgttgctaa atattgtcac tttccttgtg
     3841 gttatgtttt ggttccgtga aaccaaaaat acacgtgata atacagatac cgaagtaggg
     3901 gttgagacgc aatcaaattc ggtgtacatc actttattta aaacgatgcc cattttgttg
     3961 attatttatt tttcagcgca attgataggc caaattcccg caacggtgtg ggtgctattt
     4021 accgaaaatc gttttggatg gaatagcatg atggttggct tttcattagc gggtcttggt
     4081 cttttacact cagtattcca agcctttgtg gcaggaagaa tagccactaa atggggcgaa
     4141 aaaacggcag tactgctcga atttattgca gatagtagtg catttgcctt tttagcgttt
     4201 atatctgaag gttggttaga tttccctgtt ttaattttat tggctggtgg tgggatcgct
     4261 ttacctgcat tacagggagt gatgtctatc caaacaaaga gtcatgagca aggtgcttta
     4321 cagggattat tggtgagcct taccaatgca accggtgtta ttggcccatt actgtttact
     4381 gttatttata atcattcact accaatttgg gatggctgga tttggattat tggtttagcg
     4441 ttttactgta ttattatcct gctatcaatg accttcatgt tgacccctca agctcagggg
     4501 agtaaacagg agacaagtgc ttagttattt cgtcaccaaa tgaggcctcc aaaacttcat
     4561 ttttaatttt tgagctcgct tggactcctg tttcatgagc ggatacatat ttgaatgtat
     4621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
