LOCUS       sequence-379958-        6101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express codon-optimized human MIGA2 fragment 306-593 with a 
            C-terminal 6xhis tag (lipid transfer construct) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6101)
  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 6101)
  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: pET29-hMIGA2(306-593)-6xhis
FEATURES             Location/Qualifiers
     source          1..6101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(101..120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     311..332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(561..1376)
                     /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(561..1376)
                     /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     1284..1303
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1368..1387
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1498..2086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1987..2006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2240..2257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2272..2411
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2337..2356)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2497..2519
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2513..2704)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3513..4595)
                     /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     4557..4576
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4596..4673)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4879..4898
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4986..5005
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4986..5004
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5005..5029
                     /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             5044..5066
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5947..5964
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6017..6035)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6031..6078
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
       61 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      421 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
      481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      541 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      601 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      661 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
      721 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
      781 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
      841 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
      901 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
      961 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1021 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     1081 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1141 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1201 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     1261 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1321 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     1381 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     1441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1621 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     1681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2281 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     2341 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     2401 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2461 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2521 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     2581 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     2641 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     2701 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     2761 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     2821 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     2881 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     2941 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     3001 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     3061 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     3121 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca
     3181 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     3241 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     3301 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     3361 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     3421 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3481 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3601 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3661 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3721 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     3781 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     3841 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     3901 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     3961 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4021 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4081 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4141 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4201 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4261 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4321 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4381 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4441 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4501 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4561 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4621 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     4681 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     4741 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     4801 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     4861 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     4921 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc
     4981 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     5041 aattttgttt aactttaaga aggagatata catatgggcg actatccgat tccgctgagc
     5101 cgtccggcgg cggcgtatga ggaagcgctg caactggtga aagaaggccg tgttccgtgc
     5161 cgtaccctgc gtaccgagct gctgggttgc tacagcgacc aggattttct ggcgaagctg
     5221 cactgcgtgc gtcaagcgtt cgaaggtctg ctggaggata aaagcaacca gctgttcttt
     5281 ggcaaagtgg gtcgtcaaat ggttaccggc ctgatgacca aggcggagaa gagcccgaag
     5341 ggttttctgg agagctacga ggaaatgctg agctatgcgc tgcgtccgga aacctgggcg
     5401 accacccgtc tggagctgga aggtcgtggc gtggtttgca tgagcttctt tgacatcgtt
     5461 ctggatttta ttctgatgga cgcgttcgaa gatctggaga acccgccggc gagcgtgctg
     5521 gcggttctgc gtaaccgttg gctgagcgac agcttcaaag aaaccgcgct ggcgaccgcg
     5581 tgctggagcg tgctgaaggc gaaacgtcgt ctgctgatgg ttccggatgg tttcatcagc
     5641 cacttttaca gcgtgagcga acacgtgagc ccggttctgg cgttcggctt tctgggtccg
     5701 aaaccgcagc tggcggaagt gtgcgcgttc tttaagcacc agattgttca atacctgcgt
     5761 gacatgttcg acctggataa cgttcgttat accagcctgc cggcgctggc ggatgatatc
     5821 ctgcagctga gccgtcgtcg tagcgagatt ctgctgggtt atctgggtgt gccggcggcg
     5881 agcagcgcgg gcgttaacgg tgcgctgccg cgtgaaaacg gtccgctggg cgagctgcaa
     5941 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     6001 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     6061 gtcttgaggg gttttttgct gaaaggagga actatatccg g
//
