LOCUS       sequence-342947-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deGFP reporter plasmid with ATAAC 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-ATAAC
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     144..165
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(360..377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(531..1119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(611..630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1290..2081,2082..2150))
                     /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     1913..1932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2151..2255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2323..2341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2687..2709
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2746..2767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3007..3026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
       61 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
      121 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
      181 taactcgagc aaagcccgcc gaaaggcggg cttttctgtg tcgaccgatg cccttgagag
      241 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
      301 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
      361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      841 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      961 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1201 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1261 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     1921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2221 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2281 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2341 ccctttcgtc ttcaagaatt ctggcgaatc ctttaattaa ctgaccagcc agaaaacgac
     2401 ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca gcaagtgggg gacagcagaa
     2461 gacctgaccg ccgcagagtg gatgtttgac atggtgaaga ctatcgcacc atcagccaga
     2521 aaaccgaatt ttgctgggtg ggctaacgat atccgcctga tgcgtgaacg tgacggacgt
     2581 aaccaccgcg acatgtgtgt gcataaccgc tgggcatgct gagctaacac cgtgcgtgtt
     2641 gacaatttta cctctggcgg tgataatggt tgcagctagc aataattttg tttaacttta
     2701 agaaggagat ataccatgga gcttttcact ggcgttgttc ccatcctggt cgagctggac
     2761 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2821 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2881 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2941 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3001 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3061 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3121 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3181 ggcatcaagg tgaacttcaa gatccgccac
//
