LOCUS       sequence-66811-e         828 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 828)
  TITLE     Craig lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 828)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET15b/ECFP-GgVcl 884-1066 (Vt)
FEATURES             Location/Qualifiers
     source          1..828
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             15..37
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             56..73
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             83..100
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(104..106,107..109,110..820)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(149..170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(410..429)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     757..778
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cctctagaaa taattttgtt taactttaag aaggagatat accatgggca gcagccatca
       61 tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatggtga gcaagggcga
      121 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      181 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      241 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      301 ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      361 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      421 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      481 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      541 catcagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
      601 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      661 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
      721 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
      781 cgccgccggg atcactctcg gcatggacga gctgtacaag tcgaattc
//
