LOCUS       sequence-457802-        6440 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 6440)
  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 6440)
  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: pD474N-EGFP
FEATURES             Location/Qualifiers
     source          1..6440
                     /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"
     CDS             903..914
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(2326..2328,2329..2331,2332..3045)
                     /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(2326..2328,2329..2331,2332..3042)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2371..2392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2632..2651)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2979..3000
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3168..3289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3205..3224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3259..3278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3296..3751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3433..3454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3645..3664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3778..3882
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3884..4241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3884..4075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3907..3927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4092..4227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4276..5070
                     /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             4276..5070
                     /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(4330..4349)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4940..4959
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5258..5277)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5302..5349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5678..6266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6167..6186
                     /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 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcccacca tggctatgtc ccatcaacct
      601 ctcagctgcc tcactgaaaa ggaggacagc cccagtgaaa gcacaggaaa tggacccccc
      661 cacctggccc acccaaacct ggacacgttt accccggagg agctgctgca gcagatgaaa
      721 gagctcctga ccgagaacca ccagctgaaa gaagccatga agctaaataa tcaagccatg
      781 aaagggagat ttgaggagct ttcggcctgg acagagaaac agaaggaaga acgccagttt
      841 tttgagatac agagcaaaga agcaaaagag cgtctaatgg ccttgagtca tgagaatgag
      901 aaattgaagg aagagcttgg aaaactaaaa gggaaatcag aaaggtcatc tgaggacccc
      961 actgatgact ccaggcttcc cagggccgaa gcggagcagg aaaaggacca gctcaggacc
     1021 caggtggtga ggctacaagc agagaaggca gacctgttgg gcatcgtgtc tgaactgcag
     1081 ctcaagctga actccagcgg ctcctcagaa gattcctttg ttgaaattag gatggctgaa
     1141 ggagaagcag aagggtcagt aaaagaaatc aagcatagtc ctgggcccac gagaacagtc
     1201 tccactggca cggcattgtc taaatatagg agcagatctg cagatggggc caagaattac
     1261 ttcgaacatg aggagttaac tgtgagccag ctcctgctgt gcctaaggga agggaatcag
     1321 aaggtggaga gacttgaagt tgcactcaag gaggccaaag aaagagtttc agattttgaa
     1381 aagaaaacaa gtaatcgttc tgagattgaa acccagacag aggggagcac agagaaagag
     1441 aatgatgaag agaaaggccc ggagactgtt ggaagcgaag tggaagcact gaacctccag
     1501 gtgacatctc tgtttaagga gcttcaagag gctcatacaa aactcagcga agctgagcta
     1561 atgaagaaga gacttcaaga aaagtgtcag gcccttgaaa ggaaaaattc tgcaattcca
     1621 tcagagttga atgaaaagca agagcttgtt tatactaaca aaaagttaga gctacaagtg
     1681 gaaagcatgc tatcagaaat caaaatggaa caggctaaaa cagaggatga aaagtccaaa
     1741 ttaactgtgc tacagatgac acacaacaag cttcttcaag aacataataa tgcattgaaa
     1801 acaattgagg aactaacaag aaaagagtca gaaaaagtgg acagggcagt gctgaaggaa
     1861 ctgagtgaaa aactggaact ggcagagaag gctctggctt ccaaacagct gcaaatggat
     1921 gaaatgaagc aaaccattgc caagcaggaa gaggacctgg aaaccatgac catcctcagg
     1981 gctcagatgg aagtttactg ttctaatttt catgctgaaa gagcagcgag agagaaaatt
     2041 catgaggaaa aggagcaact ggcattgcag ctggcagttc tgctgaaaga gaatgatgct
     2101 ttcgaagacg gaggcaggca gtccttgatg gagatgcaga gtcgtcatgg ggcgagaaca
     2161 agtgactctg accagcaggc ttaccttgtt caaagaggag ctgaggacag ggactggcgg
     2221 caacagcgga atattccgat tcattcctgc cccaagtgtg gagaggttct gcctgacata
     2281 gacacgttac agattcacgt gatggattgc atcattcggg atccaatggt gagcaagggc
     2341 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2401 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gttgaccctg
     2461 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2521 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2581 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2641 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2701 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2761 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2821 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2881 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     2941 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3001 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     3061 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3121 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3181 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3241 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3301 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3361 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3421 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3481 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3541 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3601 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3661 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3721 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3901 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3961 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4081 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4141 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4201 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4261 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4321 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4381 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4441 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4501 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4561 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4621 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4681 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4741 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4801 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4861 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4921 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4981 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5041 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5101 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5161 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5221 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5281 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5341 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5401 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5461 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5521 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5821 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6361 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     6421 agcccatata tggagttccg
//
