LOCUS       sequence-329537-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deGFP reporter plasmid with GTAAT PAM upstream of p70a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3210)
  AUTHORS   Wimmer F, Englert F, Beisel CL
  TITLE     A TXTL-Based Assay to Rapidly Identify PAMs for CRISPR-Cas Systems 
            with Multi-Protein Effector Complexes.
  JOURNAL   Methods Mol Biol. 2022;2433:391-411. doi: 
            10.1007/978-1-0716-1998-8_24.
  PUBMED    34985758
REFERENCE   2  (bases 1 to 3210)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2433:391-411. doi"; pages: " 
            10.1007/978-1-0716-1998-8_24"
COMMENT     Sequence Label: pGFP-GTAAT
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(355..1146,1147..1215))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     978..997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1216..1320)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1388..1406
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             1752..1774
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1811..1832)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2072..2091)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2419..2440
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2635..2652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2806..3210,1..184))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2886..2905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
       61 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      961 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1321 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1381 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctggc gaatccttta
     1441 attaactgac cagccagaaa acgacctttc tgtggtgaaa ccggatgctg caattcagag
     1501 cgccagcaag tgggggacag cagaagacct gaccgccgca gagtggatgt ttgacatggt
     1561 gaagactatc gcaccatcag ccagaaaacc gaattttgct gggtgggcta acgatatccg
     1621 cctgatgcgt gaacgtgacg gacgtaacca ccgcgacatg tgtgtgcgta atcgctgggc
     1681 atgctgagct aacaccgtgc gtgttgacaa ttttacctct ggcggtgata atggttgcag
     1741 ctagcaataa ttttgtttaa ctttaagaag gagatatacc atggagcttt tcactggcgt
     1801 tgttcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1861 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1921 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1981 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2041 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2101 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2161 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2221 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2281 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2341 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     2401 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatctaact
     2461 cgagcaaagc ccgccgaaag gcgggctttt ctgtgtcgac cgatgccctt gagagccttc
     2521 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     2581 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     2641 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2701 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2761 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2821 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2881 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2941 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3001 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3061 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3121 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3181 agcgaggtat gtaggcggtg ctacagagtt
//
