LOCUS       sequence-99650-e        1376 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1376)
  AUTHORS   Goulas T, Cuppari A, Garcia-Castellanos R, Snipas S, Glockshuber R, 
            Arolas JL, Gomis-Ruth FX
  TITLE     The pCri System: A Vector Collection for Recombinant Protein 
            Expression and Purification.
  JOURNAL   PLoS One. 2014 Nov 11;9(11):e112643. doi: 
            10.1371/journal.pone.0112643. eCollection 2014.
  PUBMED    25386923
REFERENCE   2  (bases 1 to 1376)
  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: "PLoS One.";
            date: "2014-11-11"; volume: "9(11):e112643. doi"; pages: " 
            10.1371/journal.pone.0112643. eCollection 2014"
COMMENT     Sequence Label: pCri-12a
FEATURES             Location/Qualifiers
     source          1..1376
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    80..101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(116..121,122..139,140..146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    154..170
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     159..181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     178..194
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     178..194
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     230..292
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             join(302..304,305..1018)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(344..365)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(605..624)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     952..973
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1071..1088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
       61 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      121 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      181 gaaacagcta tgaccatgat tacgaatttc tagataacga gggcaaaaaa tgaaaaagac
      241 agctatcgcg attgcagtgg cactggccgg cttcgctacc gtagcgcagg ccatggccaa
      301 agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      361 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      421 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      481 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
      541 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      601 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      661 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      721 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
      781 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
      841 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      901 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
      961 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagg
     1021 taccggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag catcatcatc
     1081 atcatcactg actgaaaaga acgggaagat ttatgaagaa aggttttatg ttgtttactt
     1141 tgttagcggc gttttcaggc tttgctcagg ctgatgacgc ggcaattcaa caaacgttag
     1201 ccaaaatggg catcaaaagc agcgatattc agcccgcgcc tgtagctggc atgaagacag
     1261 ttctgactaa cagcggcgtg ttgtacatca ccgatgatgg taaacatatc attcaggggc
     1321 caatgtatga cgttagtggc acggctccgg tcaatgtcac caataaaatg ctgtta
//
