LOCUS       sequence-339525-        5480 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses a human myelin oligodendrocyte glycoprotein isoform beta1 
            - EGFP fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5480)
  AUTHORS   Schanda K, Peschl P, Lerch M, Seebacher B, Mindorf S, Ritter N, 
            Probst M, Hegen H, Di Pauli F, Wendel EM, Lechner C, Baumann M, 
            Mariotto S, Ferrari S, Saiz A, Farrell M, Leite MIS, Irani SR, 
            Palace J, Lutterotti A, Kumpfel T, Vukusic S, Marignier R, Waters P,
            Rostasy K, Berger T, Probst C, Hoftberger R, Reindl M
  TITLE     Differential Binding of Autoantibodies to MOG Isoforms in 
            Inflammatory Demyelinating Diseases.
  JOURNAL   Neurol Neuroimmunol Neuroinflamm. 2021 Jun 15;8(5). pii: 8/5/e1027. 
            doi: 10.1212/NXI.0000000000001027. Print 2021 Jul.
  PUBMED    34131067
REFERENCE   2  (bases 1 to 5480)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neurol 
            Neuroimmunol Neuroinflamm."; date: "2021-06-15"; volume: "8(5). pii:
            8/5/e1027. doi"; pages: " 10.1212/NXI.0000000000001027. Print 2021 
            Jul"
COMMENT     Sequence Label: pEGFP-N1 hMOG-beta1-EGFP
FEATURES             Location/Qualifiers
     source          1..5480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        166..357
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     314..334
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(359..463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      490..945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(577..596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     787..808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    952..1073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(963..982)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1017..1036
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1196..1909,1910..1912,1913..1915))
                     /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             complement(join(1199..1909,1910..1912,1913..1915))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1241..1262)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1590..1609
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1849..1870
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1912..1921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2773..2976)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2802..2822)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2977..3280
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3455..4043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3535..3554)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4372..4419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4444..4463
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4651..5445)
                     /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             complement(4651..5445)
                     /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(4762..4781)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5372..5391
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5480,1..357))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
       61 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      121 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      181 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      241 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      301 ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac
      361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      481 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      541 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      601 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      661 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      721 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      781 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
      841 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
      901 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcc ttaagataca
      961 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1021 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1081 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1141 agtaaaacct ctacaaatgt ggtatggctg attatgatct agagtcgcgg ccgctttact
     1201 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     1261 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt
     1321 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     1381 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     1441 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     1501 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     1561 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     1621 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     1681 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     1741 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     1801 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     1861 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     1921 cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcggcca gagagggctt
     1981 ccaggtggaa gagtagctct tcaaggaatt gccctgctag tcttcgatgt agccagttgt
     2041 agcagatgat caaggcaacc aagggtccaa gaaccggcac aattacaaac agggttatct
     2101 tccagcaggg caccctcaga aagtggggat caaaagtccg gtggagattc tctatctctg
     2161 ctcgaagttt tcctctcagt ctgtactgca ggcagaggaa gacgaggcca acagtgatct
     2221 gcaggaggag cacaggcagc accgcgagga gaaccagcac tccagggctc acccagtaga
     2281 aaggatcttc tactttcaat tccattgctg cctcctcttg gtaagaatga tctcggaaga
     2341 agcaggtgaa acctccttca tctgagaacc ttacattccg gatcctgaga gtcacctttc
     2401 cctcaccaat agcatctttc agcagctctg tccggccccg atattcaggt gcctggtctc
     2461 catcttggtc cttgccattt ctgtagagat gaaccaccct agagaagggg gggcggtacc
     2521 accccacctc catgcctgta gcgttcttcc caggagatat gcgacatggc aattccactt
     2581 catccccgac cagagcccgg atagggtgtc ttggtcctat cactctgaac tgccctgcat
     2641 agctggaaga cacttggagg aggaggagga ggaggaagga gcagaggcag ctgggcagag
     2701 agggtcttga taagcttgcc atctcgagat ctgagtccgg tagcgctagc ggatctgacg
     2761 gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     2821 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     2881 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     2941 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     3001 gtagatatac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     3061 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     3121 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     3181 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     3241 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     3301 atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt
     3361 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3421 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3481 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3541 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3601 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3661 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3721 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3781 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3841 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3901 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3961 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4021 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4081 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4141 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac
     4201 ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg
     4261 aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc
     4321 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
     4381 acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     4441 gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca
     4501 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     4561 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     4621 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     4681 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     4741 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     4801 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     4861 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc
     4921 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     4981 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     5041 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     5101 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     5161 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     5221 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc
     5281 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     5341 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     5401 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     5461 tcctgtctct tgatcgatct
//
