LOCUS       sequence-287557-        6439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutated OPTN was inserted into pEGFP-N1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6439)
  AUTHORS   Turturro S, Shen X, Shyam R, Yue BY, Ying H
  TITLE     Effects of mutations and deletions in the human optineurin gene.
  JOURNAL   Springerplus. 2014 Feb 19;3:99. doi: 10.1186/2193-1801-3-99. 
            eCollection 2014.
  PUBMED    24683533
REFERENCE   2  (bases 1 to 6439)
  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: 
            "Springerplus."; date: "2014-02-19"; volume: "3:99. doi"; pages: " 
            10.1186/2193-1801-3-99. eCollection 2014"
COMMENT     Sequence Label: pE478G-EGFP
FEATURES             Location/Qualifiers
     source          1..6439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             621..644
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             962..973
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(2385..2387,2388..2390,2391..3104)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2385..2387,2388..2390,2391..3101)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2430..2451)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2691..2710)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3038..3059
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3227..3348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3264..3283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3318..3337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3355..3810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3492..3513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3704..3723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3837..3941
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3943..4300
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3943..4134
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3966..3986)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4151..4286
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4335..5129
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4335..5129
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4389..4408)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4999..5018
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5317..5336)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5361..5408
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5737..6325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6226..6245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcctcg
      601 agacgcgtga ttccaccatg gattacaagg atgacgacga taagatgtcc catcaacctc
      661 tcagctgcct cactgaaaag gaggacagcc ccagtgaaag cacaggaaat ggaccccccc
      721 acctggccca cccaaacctg gacacgttta ccccggagga gctgctgcag cagatgaaag
      781 agctcctgac cgagaaccac cagttgaaag aagccatgaa gctaaataat caagccatga
      841 aagggagatt tgaggagctt tcggcctgga cagagaaaca gaaggaagaa cgccagtttt
      901 ttgagataca gagcaaagaa gcaaaagagc gtctaatggc cttgagtcat gagaatgaga
      961 aattgaagga agagcttgga aaactaaaag ggaaatcaga aaggtcatct gaggacccca
     1021 ctgatgactc caggcttccc agggccgaag cggagcagga aaaggaccag ctcaggaccc
     1081 aggtggtgag gctacaagca gagaaggcag acctgttggg catcgtgtct gaactgcagc
     1141 tcaagctgaa ctccagcggc tcctcagaag attcctttgt tgaaattagg atggctgaag
     1201 gagaagcaga agggtcagta aaagaaatca agcatagtcc tgggcccacg agaacagtct
     1261 ccactggcac ggcattgtct aaatatagga gcagatctgc agatggggcc aagaattact
     1321 tcgaacatga ggagttaact gtgagccagc tcctgctgtg cctaagggaa gggaatcaga
     1381 aggtggagag acttgaagtt gcactcaagg aggccaaaga aagagtttca gattttgaaa
     1441 agaaaacaag taatcgttct gagattgaaa cccagacaga ggggagcaca gagaaagaga
     1501 atgatgaaga gaaaggcccg gagactgttg gaagcgaagt ggaagcactg aacctccagg
     1561 tgacatctct gtttaaggag cttcaagagg ctcatacaaa actcagcgaa gctgagctaa
     1621 tgaagaagag acttcaagaa aagtgtcagg cccttgaaag gaaaaattct gcaattccat
     1681 cagagttgaa tgaaaagcaa gagcttgttt atactaacaa aaagttagag ctacaagtgg
     1741 aaagcatgct atcagaaatc aaaatggaac aggctaaaac agaggatgaa aagtccaaat
     1801 taactgtgct acagatgaca cacaacaagc ttcttcaaga acataataat gcattgaaaa
     1861 caattgagga actaacaaga aaagagtcag aaaaagtgga cagggcagtg ctgaaggaac
     1921 tgagtgaaaa actggaactg gcagagaagg ctctggcttc caaacagctg caaatggatg
     1981 aaatgaagca aaccattgcc aagcaggaag aggacctgga aaccatgacc atcctcaggg
     2041 ctcagatgga agtttactgt tctgattttc atgctggaag agcagcgaga gagaaaattc
     2101 atgaggaaaa ggagcaactg gcattgcagc tggcagttct gctgaaagag aatgatgctt
     2161 tcgaagacgg aggcaggcag tccttgatgg agatgcagag tcgtcatggg gcgagaacaa
     2221 gtgactctga ccagcaggct taccttgttc aaagaggagc tgaggacagg gactggcggc
     2281 aacagcggaa tattccgatt cattcctgcc ccaagtgtgg agaggttctg cctgacatag
     2341 acacgttaca gattcacgtg atggattgca tcattcggga tccaatggtg agcaagggcg
     2401 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2461 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2521 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2581 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2641 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2701 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2761 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2821 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2881 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2941 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3001 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3061 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     3121 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3181 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3241 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3301 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3361 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3421 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3481 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3541 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3601 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3661 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3721 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3781 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3841 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3901 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3961 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4021 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4141 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4201 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4261 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4321 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4381 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4441 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4501 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4561 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4621 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4681 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4741 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4801 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4861 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4921 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4981 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5041 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5101 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5161 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5221 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5281 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5341 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5401 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5461 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5521 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5581 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5641 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5701 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5761 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5821 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5881 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5941 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6001 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6061 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6121 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6181 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6241 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6301 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6361 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6421 ccgtattacc gccatgcat
//
