LOCUS       sequence-123824-        3670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial constitutive gRNA expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3670)
  TITLE     Bacterial constitutive gRNA expression plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3670)
  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            "Unpublished"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: pFG001
FEATURES             Location/Qualifiers
     source          1..3670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(53..121,122..913)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(271..290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1070..1145
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     repeat_region   1201..1236
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   1267..1302
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_feature    complement(1303..1434)
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     CDS             complement(1971..2921)
                     /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      2969..3191
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
       61 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
      181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      241 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
      301 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
      361 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
      421 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
      481 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
      541 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
      601 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
      661 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
      721 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
      781 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
      841 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
      901 taagcattgg taacaggtcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg
      961 gtttttgctt ttggaggggc agaaagatga atgactgtct cataaaaaat ttatttgctt
     1021 tcaggaaaat ttttctgtat aatagattca tgctcgatga gtttttctag ttttagagct
     1081 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     1141 ggtgctttta aaaaaaaaac cccgcgcggg gttttttttt acaagtctca gtgtgctgaa
     1201 gttttgggac cattcaaaac agcatagctc taaaacgaga cctttgagct tccgagactg
     1261 gtctcagttt tgggaccatt caaaacagca tagctctaaa acctcgtaga ctatttttgt
     1321 ctaaaaaatt tcgtaatcgc actatttgtc tcagctagac ttcagtcttg aaaagcccct
     1381 gtattactgc atttattaag agtattatac catattttta gttattaaga aataatcttc
     1441 atctaaaata tactacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt
     1501 gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc
     1561 agaattgcca tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct
     1621 ttgattcgat aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa
     1681 caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg
     1741 ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag
     1801 ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     1861 catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     1921 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     1981 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     2041 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     2101 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     2161 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     2221 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     2281 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     2341 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     2401 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     2461 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     2521 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     2581 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     2641 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     2701 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     2761 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     2821 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     2881 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     2941 tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc
     3001 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     3061 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     3121 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     3181 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     3241 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     3301 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     3361 ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca
     3421 ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt
     3481 cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc
     3541 tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca
     3601 ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg
     3661 tcttactgtc
//
