LOCUS       sequence-379955-        7556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express codon-optimized human MIGA2 fragment 170-593 in Expi293 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7556)
  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 7556)
  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: pCMV10-3xFLAG-prs-hMIGA2 (170-593)
FEATURES             Location/Qualifiers
     source          1..7556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..155
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     147..164
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     148..164
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        325..704
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        705..908
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     859..879
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             938..1003
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1004..1027
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2322..2944
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2521..2540)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2968..2988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2986..3283
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3337..4131
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3391..3410)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4001..4020
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4783..4917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4820..4839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4874..4893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4922..4941)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4923..4941)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4955..4971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4955..4971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4968..4990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4979..4995
                     /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)."
     promoter        complement(join(5003..5009,5010..5027,5028..5033))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5186..5203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5357..5945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5437..5456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6116..6907,6908..6976))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6739..6758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6977..7081)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7108..7536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7195..7214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7405..7426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
       61 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      121 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgatctata
      181 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
      241 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
      301 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
      361 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      421 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      481 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      541 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
      601 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
      661 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      721 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      781 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      841 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      901 gcagagctcg tttagtgaac cgtcagaatt aaccatggac tacaaagacc atgacggtga
      961 ttataaagat catgacatcg attacaagga tgacgatgac aagctggaag ttctgttcca
     1021 ggggcccctt gcggccgccg atggcaacgc ggaaagcctg tacatgcagg gtatggagct
     1081 gtttgaggaa gcgctgcaga agtgggagca agcgctgagc gttggtcaac gtggcgacag
     1141 cggtagcacc ccgatgccgc gtgatggtct gcgtaacccg gaaaccgcga gcgagccgct
     1201 gagcgagccg gaaagccagc gtaaagagtt cgcggaaaag ctggagagcc tgctgcaccg
     1261 tgcgtatcac ctgcaagagg aattcggcag cacctttccg gcggacagca tgctgctgga
     1321 tctggaacgt accctgatgc tgccgctgac cgagggtagc ctgcgtctgc gtgcggacga
     1381 tgaagacagc ctgaccagcg aggatagctt ctttagcgcg accgaactgt tcgagagcct
     1441 gcagaccggc gactatccga ttccgctgag ccgtccggcg gcggcgtatg aggaagcgct
     1501 gcaactggtg aaagaaggcc gtgttccgtg ccgtaccctg cgtaccgagc tgctgggttg
     1561 ctacagcgac caggattttc tggcgaagct gcactgcgtg cgtcaagcgt tcgaaggtct
     1621 gctggaggat aaaagcaacc agctgttctt tggcaaagtg ggtcgtcaaa tggttaccgg
     1681 cctgatgacc aaggcggaga agagcccgaa gggttttctg gagagctacg aggaaatgct
     1741 gagctatgcg ctgcgtccgg aaacctgggc gaccacccgt ctggagctgg aaggtcgtgg
     1801 cgtggtttgc atgagcttct ttgacatcgt tctggatttt attctgatgg acgcgttcga
     1861 agatctggag aacccgccgg cgagcgtgct ggcggttctg cgtaaccgtt ggctgagcga
     1921 cagcttcaaa gaaaccgcgc tggcgaccgc gtgctggagc gtgctgaagg cgaaacgtcg
     1981 tctgctgatg gttccggatg gtttcatcag ccacttttac agcgtgagcg aacacgtgag
     2041 cccggttctg gcgttcggct ttctgggtcc gaaaccgcag ctggcggaag tgtgcgcgtt
     2101 ctttaagcac cagattgttc aatacctgcg tgacatgttc gacctggata acgttcgtta
     2161 taccagcctg ccggcgctgg cggatgatat cctgcagctg agccgtcgtc gtagcgagat
     2221 tctgctgggt tatctgggtg tgccggcggc gagcagcgcg ggcgttaacg gtgcgctgcc
     2281 gcgtgaaaac ggtccgctgg gcgagctgca ataaggatcc cgggtggcat ccctgtgacc
     2341 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2401 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2461 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2521 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2581 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     2641 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     2701 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     2761 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     2821 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     2881 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     2941 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3121 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3181 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3241 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggaatt gatcagcttg
     3301 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3361 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3421 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3481 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3541 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3601 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3661 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3721 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3781 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3841 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3901 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3961 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4021 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4081 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4141 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4201 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4261 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg
     4321 cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca
     4381 aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc
     4441 aggagtgggg aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga
     4501 tacagaacga attgcttgca ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt
     4561 cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc gagctgctca ccacccactc
     4621 gccaagctgg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccgatc
     4681 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4741 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4801 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4861 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat
     4921 tccctatagt gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     4981 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5041 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5101 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5161 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5221 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5281 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5341 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5401 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5461 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5521 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5581 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5641 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5701 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5761 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5821 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5881 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5941 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6001 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6061 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6121 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6181 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6241 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6301 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6361 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6421 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6481 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6541 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6601 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6661 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6721 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6781 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6841 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6901 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6961 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7021 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7081 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     7141 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     7201 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7261 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7321 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7381 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7441 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7501 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgctt
//
