LOCUS       sequence-342943-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deGFP reporter plasmid with CAAAG 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-CAAAG
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     194..215
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(410..427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(581..1169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(661..680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1340..2131,2132..2200))
                     /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     1963..1982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2201..2305)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2373..2391
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2737..2759
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2796..2817)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3057..3076)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
       61 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      121 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
      181 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc taactcgagc
      241 aaagcccgcc gaaaggcggg cttttctgtg tcgaccgatg cccttgagag ccttcaaccc
      301 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
      361 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
      421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1021 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1921 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     1981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2341 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2401 ttcaagaatt ctggcgaatc ctttaattaa ctgaccagcc agaaaacgac ctttctgtgg
     2461 tgaaaccgga tgctgcaatt cagagcgcca gcaagtgggg gacagcagaa gacctgaccg
     2521 ccgcagagtg gatgtttgac atggtgaaga ctatcgcacc atcagccaga aaaccgaatt
     2581 ttgctgggtg ggctaacgat atccgcctga tgcgtgaacg tgacggacgt aaccaccgcg
     2641 acatgtgtgt gccaaagcgc tgggcatgct gagctaacac cgtgcgtgtt gacaatttta
     2701 cctctggcgg tgataatggt tgcagctagc aataattttg tttaacttta agaaggagat
     2761 ataccatgga gcttttcact ggcgttgttc ccatcctggt cgagctggac ggcgacgtaa
     2821 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2881 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2941 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     3001 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3061 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3121 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3181 acaactacaa cagccacaac gtctatatca
//
