LOCUS       sequence-457800-        6557 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 6557)
  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 6557)
  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: pR96L-EGFP
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1327..1350
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1668..1679
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3082..3108
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3285..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3322..3341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3376..3395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3413..3868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3571)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3762..3781
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3895..3999
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4001..4358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4001..4192
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4024..4044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4209..4344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4393..5187
                     /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             4393..5187
                     /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(4447..4466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5057..5076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5375..5394)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5419..5466
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6284..6303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgccaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga atttcgagac gcgtgattcc
     1321 accatggatt acaaggatga cgacgataag atgtcccatc aacctctcag ctgcctcact
     1381 gaaaaggagg acagccccag tgaaagcaca ggaaatggac ccccccacct ggcccaccca
     1441 aacctggaca cgtttacccc ggaggagctg ctgcagcaga tgaaagagct cctgaccgag
     1501 aaccaccagc tgaaagaagc catgaagcta aataatcaag ccatgaaagg gagatttgag
     1561 gagctttcgg cctggacaga gaaacagaag gaagaacgcc agttttttga gatacagagc
     1621 aaagaagcaa aagagcttct aatggccttg agtcatgaga atgagaaatt gaaggaagag
     1681 cttggaaaac taaaagggaa atcagaaagg tcatctgagg accccactga tgactccagg
     1741 cttcccaggg ccgaagcgga gcaggaaaag gaccagctca ggacccaggt ggtgaggcta
     1801 caagcagaga aggcagacct gttgggcatc gtgtctgaac tgcagctcaa gctgaactcc
     1861 agcggctcct cagaagattc ctttgttgaa attaggatgg ctgaaggaga ggcagaaggg
     1921 tcagtaaaag aaatcaagca tagtcctggg cccacgagaa cagtctccac tggcacggca
     1981 ttgtctaaat ataggagcag atctgcagat ggggccaaga attacttcga acatgaggag
     2041 ttaactgtga gccagctcct gctgtgccta agggaaggga atcagaaggt ggagagactt
     2101 gaagttgcac tcaaggaggc caaagaaaga gtttcagatt ttgaaaagaa aacaagtaat
     2161 cgttctgaga ttgaaaccca gacagagggg agcacagaga aagagaatga tgaagagaaa
     2221 ggcccggaga ctgttggaag cgaagtggaa gcactgaacc tccaggtgac atctctgttt
     2281 aaggagcttc aagaggctca tacaaaactc agcgaagctg agctaatgaa gaagagactt
     2341 caagaaaagt gtcaggccct tgaaaggaaa aattctgcaa ttccatcaga gttgaatgaa
     2401 aagcaagagc ttgtttatac taacaaaaag ttagagctac aagtggaaag catgctatca
     2461 gaaatcaaaa tggaacaggc taaaacagag gatgaaaagt ccaaattaac tgtgctacag
     2521 atgacacaca acaagcttct tcaagaacat aataatgcat tgaaaacaat tgaggaacta
     2581 acaagaaaag agtcagaaaa agtggacagg gcagtgctga aggaactgag tgaaaaactg
     2641 gaactggcag agaaggctct ggcttccaaa cagctgcaaa tggatgaaat gaagcaaacc
     2701 attgccaagc aggaagagga cctggaaacc atgaccatcc tcagggctca gatggaagtt
     2761 tactgttctg attttcatgc tgaaagagca gcgagagaga aaattcatga ggaaaaggag
     2821 caactggcat tgcagctggc agttctgctg aaagagaatg atgctttcga agacggaggc
     2881 aggcagtcct tgatggagat gcagagtcgt catggggcga gaacaagtga ctctgaccag
     2941 caggcttacc ttgttcaaag aggagctgag gacagggact ggcggcaaca gcggaatatt
     3001 ccgattcatt cctgccccaa gtgtggagag gttctgcctg acatagacac gttacagatt
     3061 cacgtgatgg attgcatcat ttacccatac gacgtcccag actacgctta gcaaatcttt
     3121 cccgggggta ccgtcgagac ggtaccgcgg gcccgggatc caccggatct agataactga
     3181 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3241 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3421 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3481 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3541 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3601 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3661 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3721 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3781 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3841 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3901 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3961 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4021 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4201 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4261 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4321 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4381 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4561 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4921 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5221 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5281 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5341 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5401 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5461 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5521 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5581 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5641 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5701 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     6541 ccatatatgg agttccg
//
