LOCUS       sequence-342948-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deGFP reporter plasmid with unique PacI restriction enzyme 
            recognition site.
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-PacI
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(60..81)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(321..340)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     668..689
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(884..901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1055..1643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1135..1154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1814..2605,2606..2674))
                     /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     2437..2456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2675..2779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2847..2865
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacca tggagctttt cactggcgtt gttcccatcc
       61 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      121 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      181 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      241 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      301 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      361 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      421 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      481 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
      541 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
      601 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
      661 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatctaactc gagcaaagcc
      721 cgccgaaagg cgggcttttc tgtgtcgacc gatgcccttg agagccttca acccagtcag
      781 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
      841 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
      901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1861 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1921 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1981 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2041 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2101 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2161 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2221 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2281 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2341 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2401 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     2461 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2521 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2581 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2641 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2701 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2761 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2821 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     2881 aattctggcg aatcctttaa ttaactgacc agccagaaaa cgacctttct gtggtgaaac
     2941 cggatgctgc aattcagagc gccagcaagt gggggacagc agaagacctg accgccgcag
     3001 agtggatgtt tgacatggtg aagactatcg caccatcagc cagaaaaccg aattttgctg
     3061 ggtgggctaa cgatatccgc ctgatgcgtg aacgtgacgg acgtaaccac cgcgacatgt
     3121 gtgtgctgtt ccgctgggca tgctgagcta acaccgtgcg tgttgacaat tttacctctg
     3181 gcggtgataa tggttgcagc tagcaataat
//
