LOCUS       sequence-379954-        6780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express C.elegans MIGA fragment 106-496 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6780)
  AUTHORS   Hong Z, Adlakha J, Wan N, Guinn E, Giska F, Gupta K, Melia TJ, 
            Reinisch KM
  TITLE     Mitoguardin-2-mediated lipid transfer preserves mitochondrial 
            morphology and lipid droplet formation.
  JOURNAL   J Cell Biol. 2022 Dec 5;221(12):e202207022. doi: 
            10.1083/jcb.202207022. Epub 2022 Oct 25.
  PUBMED    36282247
REFERENCE   2  (bases 1 to 6780)
  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 Cell 
            Biol."; date: "2022-12-5"; volume: "221(12):e202207022. doi"; pages:
            " 10.1083/jcb.202207022. Epub 2022 Oct 25"
COMMENT     Sequence Label: pET28-6xhis-sumo-CeMIGA(106-496)
FEATURES             Location/Qualifiers
     source          1..6780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     559..578
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(598..675)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     881..900
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     984..1003
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        984..1002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1003..1027
                     /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             1042..1064
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1083..1100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1110..1127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1140..1433
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             2628..2645
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2698..2716)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2712..2759
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2796..3251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2883..2902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3093..3114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3343..4158)
                     /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(3343..4158)
                     /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     4066..4085
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4150..4169
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4280..4868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4769..4788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5022..5039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5054..5193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5119..5138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5279..5301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5295..5486)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6295..6780,1..597))
                     /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"
ORIGIN
        1 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
       61 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
      121 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
      181 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
      241 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
      301 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
      361 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
      421 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
      481 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
      541 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
      601 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
      661 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
      721 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
      781 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
      841 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
      901 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
      961 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     1021 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
     1081 gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat atggctagca
     1141 tgtcggactc agaagtcaat caagaagcta agccagaggt caagccagaa gtcaagcctg
     1201 agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc aagatcaaaa
     1261 agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt aaggaaatgg
     1321 actccttaag attcttgtac gacggtatta gaattcaagc tgatcagacc cctgaagatt
     1381 tggacatgga ggataacgat attattgagg ctcacagaga acagattggt ggatccagta
     1441 tgaatccaat tgaagcagac atcacaagtg gtcaagaact ttgcacagaa ctacgaaaga
     1501 ctatagaaaa agtccatcat aatctagaaa tggttaaaaa taggagtagc aaagacatgg
     1561 agagatcaat aaaaattgaa ggcattctga aaggattgaa acaagttgaa gatgaaattg
     1621 tccttctggt ccctcaaatg gaagacttcc gagatgataa catggagttc tattctgtca
     1681 gtggtggttc aggctacgcc ggaagcgttc gaacaggaag atcacgaact ctttctgtcc
     1741 tttccgatga ttcattccgt tctgcagtag aagaatttgc atgtgacatt gacgatatcg
     1801 attttgttag tgatgcagcg aatttggata aaaatgaact gcgatttttg gatgaaggaa
     1861 tgcaagcggc gttaaatgga gaagtaaagt atagaaaaag tcgaatggaa ttctgcaagt
     1921 gtgacagtga aacagacttt gctgccaagt tatactgtgt taggcaagct ctcacgaacg
     1981 ctttgaaaga cgagcacaaa cgtgtatggc ttgcgaaatg tggccgaact ctcctcgctg
     2041 attttattcg tcacacgaaa caggatcctg tgaagttctt taatgcctac gatgagatgc
     2101 tggaatatgt atcaaatgac cgtaatgaag agcagctcag acaagacgtc gaaggccgag
     2161 gtgtttgtga aactggcttt tatgatgtcg caatcgattt tattatcctc gatgcatttg
     2221 aagacttaaa atcaccacct tctgcagttt attccgtcac caagaattat tttatgtcga
     2281 tgtccatgaa gtactctact ctcaacacta tcatctggtc tatcattaaa tcaaagcgtc
     2341 aaagactgaa aaatcctgat ggttttattg caaagttcta caatatatcg gaaactgtga
     2401 tgccggccat cactctcggc tttttgggaa ctgatgaacg cctcggagaa ttgtgtcaat
     2461 attttaagga acaagttgta cagttcgtac tagatgtttt caatacacag aaggtttgct
     2521 atcgatcgtt agaggagatg tcagaagatg tttggattgt gatgcgtaat cgattagaag
     2581 cagttcagac aagaatgagc aacgaactat aagcggccgc actcgagcac caccaccacc
     2641 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     2701 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2761 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     2821 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2881 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2941 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3001 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3061 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3121 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3181 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3241 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3301 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     3361 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     3421 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     3481 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     3541 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     3601 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     3661 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     3721 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     3781 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     3841 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     3901 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     3961 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     4021 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     4081 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     4141 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     4201 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4261 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4321 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4381 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4801 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4861 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4921 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4981 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5041 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5101 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     5161 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     5221 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5281 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     5341 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     5401 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     5461 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     5521 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     5581 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     5641 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     5701 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     5761 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     5821 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     5881 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     5941 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     6001 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     6061 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     6121 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     6181 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     6241 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     6301 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6361 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     6421 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6481 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6541 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     6601 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     6661 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     6721 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
//
