LOCUS       sequence-449491-        5618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the soluble domain of Myelin Oligodendrocyte Glycoprotein 
            (rhMOG) in SHuffle E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5618)
  AUTHORS   Wu W, Gupta S, Sagan SA, Moseley CE, Zamvil SS, Pak JE
  TITLE     High-yield production of recombinant human myelin oligodendrocyte 
            glycoprotein in SHuffle bacteria without a refolding step.
  JOURNAL   J Immunol Methods. 2024 Oct 13:113764. doi: 
            10.1016/j.jim.2024.113764.
  PUBMED    39406335
REFERENCE   2  (bases 1 to 5618)
  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: "J Immunol 
            Methods. 2024 Oct 13:113764. doi: 10.1016/j.jim.2024.113764."
COMMENT     Sequence Label: pET29b-rhMOG
FEATURES             Location/Qualifiers
     source          1..5618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             361..378
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             388..405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(458..476)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      472..519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      556..1011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(643..662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     853..874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1103..1918)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1103..1918)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1826..1845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1910..1929
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2040..2628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2529..2548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2782..2799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2814..2953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2879..2898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3039..3061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3055..3246)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4055..5137)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5099..5118
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5138..5215)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5421..5440
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5528..5547
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5528..5546
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5547..5571
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5586..5608
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ggacaattta gagtaatagg gcccaggcat ccgattcgtg cgctggttgg cgacgaggtg
       61 gagttaccgt gtcgtatctc cccgggtaag aacgccaccg gtatggaggt aggctggtat
      121 agaccaccgt tcagccgtgt tgttcatctg taccgcaatg gcaaagacca agatggtgac
      181 caggctccgg aataccgcgg tcgtaccgaa ttgctgaagg acgcgattgg tgaaggcaaa
      241 gtcaccctgc gtatccgcaa cgtgcgtttt tctgatgagg gcggttttac gtgcttcttt
      301 cgtgatcata gctatcagga ggaagcggca atggagttga aggtggaaga tccgttctac
      361 catcaccacc accaccacta actcgagcac caccaccacc accactgaga tccggctgct
      421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      541 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      601 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      661 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      721 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      781 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      841 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
      901 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
      961 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1081 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     1141 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1201 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1261 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1321 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     1381 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     1441 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     1501 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1561 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     1621 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1681 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1741 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     1801 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1861 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     1921 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     1981 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2041 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2101 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2161 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2221 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2281 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2341 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2401 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2461 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2521 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2581 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2641 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2701 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2761 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2821 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     2881 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2941 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3061 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3121 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3181 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3241 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3301 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3361 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3421 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     3481 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3541 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3601 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3661 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3721 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3781 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3841 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3901 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3961 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     4021 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4141 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     4201 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4261 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     4321 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4381 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4441 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4501 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4561 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4621 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4681 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4741 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4801 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4861 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4921 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4981 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     5041 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     5101 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     5161 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     5221 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     5281 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     5341 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     5401 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     5461 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc
     5521 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     5581 ataattttgt ttaactttaa gaaggagata tacatatg
//
