LOCUS       sequence-342949-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deGFP reporter plasmid with CAATG 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-CAATG
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(406..423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(577..1165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(657..676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1336..2127,2128..2196))
                     /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     1959..1978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2197..2301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2369..2387
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2733..2755
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2792..2813)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3053..3072)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
       61 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      121 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
      181 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatctaac tcgagcaaag
      241 cccgccgaaa ggcgggcttt tctgtgtcga ccgatgccct tgagagcctt caacccagtc
      301 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
      361 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
      421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1921 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     1981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2341 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     2401 agaattctgg cgaatccttt aattaactga ccagccagaa aacgaccttt ctgtggtgaa
     2461 accggatgct gcaattcaga gcgccagcaa gtgggggaca gcagaagacc tgaccgccgc
     2521 agagtggatg tttgacatgg tgaagactat cgcaccatca gccagaaaac cgaattttgc
     2581 tgggtgggct aacgatatcc gcctgatgcg tgaacgtgac ggacgtaacc accgcgacat
     2641 gtgtgtgcca atgcgctggg catgctgagc taacaccgtg cgtgttgaca attttacctc
     2701 tggcggtgat aatggttgca gctagcaata attttgttta actttaagaa ggagatatac
     2761 catggagctt ttcactggcg ttgttcccat cctggtcgag ctggacggcg acgtaaacgg
     2821 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2881 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2941 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3001 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3061 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3121 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3181 ctacaacagc cacaacgtct atatcatggc
//
