LOCUS       sequence-490711-        1926 bp    DNA     linear   UNA 22-JAN-2026
DEFINITION  Expresses ALFA-bRho-EGFP in mammalian cells. Modified from Addgene 
            plasmid #45399.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1926)
  AUTHORS   Hekmatara M, Thompson SL, Haggerty KN, Hagen S, Brothers BA, Daniels
            B, Luxardi G, Moshiri A, Deng WT, Robichaux MA
  TITLE     Super-resolution microscopy reveals a Rab6a-dependent trafficking 
            hub for rhodopsin at the mammalian rod photoreceptor Golgi.
  JOURNAL   bioRxiv [Preprint]. 2025 Sep 25:2025.09.24.678292. doi: 
            10.1101/2025.09.24.678292.
  PUBMED    41040232
REFERENCE   2  (bases 1 to 1926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 22, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Sep 25:2025.09.24.678292. doi: 
            10.1101/2025.09.24.678292."
COMMENT     Sequence Label: pcDNA3.1-N-ALFA-bRho-EGFP
FEATURES             Location/Qualifiers
     source          1..1926
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             73..111
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             124..162
                     /codon_start=1
                     /product="segment of bovine rhodopsin containing a signal 
                     for N-linked glycosylation (Favaloro et al., 2008)"
                     /label=opsin tag
                     /note="The asparagine in the NKT tripeptide will be 
                     glycosylated if this sequence is exposed to the ER lumen."
                     /translation="GPNFYVPFSNKTG"
     CDS             join(1153..1155,1156..1158,1159..1869)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1153..1155,1156..1158,1159..1869)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1198..1219)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1459..1478)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1806..1827
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gtntnaactt aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg
       61 aattccacca tgagccgtct ggaagaagaa ctgcgccgcc gtctgaccga aaacggtacc
      121 gaaggcccaa acttctacgt tcctttctcc aacaagacgg gcgtggtgcg cagcccgttc
      181 gaggctccgc agtactacct ggcggagccc tggcagttct ccatgctggc cgcctacatg
      241 ttcctgctga tcatgcttgg cttcccgatc aacttcctca cgctgtacgt cacagtccag
      301 cacaagaagc ttcgcacacc gctcaactac atcctgctca acctggccgt ggcagatctc
      361 ttcatggtct tcggtggctt caccaccacc ctctacacct ctctccatgg gtacttcgtc
      421 tttgggccga cgggctgcaa cctcgagggc ttctttgcca ccctgggcgg tgaaattgca
      481 ctgtggtctc tggtagtact ggcgatcgag cggtacgtgg tggtgtgcaa gcccatgagc
      541 aacttccgct tcggtgagaa ccacgccatc atgggcgtcg ccttcacctg ggtcatggct
      601 ctggcctgtg cggccccgcc gctcgtcggc tggtctagat acatcccgga gggcatgcag
      661 tgctcgtgcg ggatcgatta ctacacgccg cacgaggaga ccaacaatga gtcgttcgtc
      721 atctacatgt tcgtggtcca cttcatcatc ccgctgattg tcatcttctt ctgctatggc
      781 cagctggtgt tcaccgtcaa ggaggctgca gcccagcagc aggagagcgc caccactcag
      841 aaggccgaga aggaggtcac gcgtatggtt atcatcatgg tcatcgcttt cctaatctgc
      901 tggctgccat atgctggtgt ggcgttctac atcttcaccc atcagggctc tgactttggg
      961 cccatcttca tgaccatccc ggctttcttt gccaagacgt ctgccgtcta caacccggtc
     1021 atctacatca tgatgaacaa gcagttccgg aactgcatgg tcaccactct ctgctgtggc
     1081 aagaacccgc tgggtgacga cgaggcgtcg accaccgtct ccaagacaga gaccagccaa
     1141 gtggcgcctg ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1201 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1261 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1321 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1381 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1441 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1501 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1561 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1621 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1681 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1741 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1801 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1861 ctgtacaagg agaccagcca agtggcgcct gcctaagcgg ccgctcgagt ctagagnccc
     1921 cnnnnc
//
