LOCUS       sequence-379957-        7649 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express codon-optimized human MIGA2 fragment 170-593 with a 
            GS-linker and 6xhis tag at C-terminus (lipid transfer construct) in 
            Expi293 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7649)
  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 7649)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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)-25GS-6xhis
FEATURES             Location/Qualifiers
     source          1..7649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             997..1020
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2380..2397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2408..3030
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2607..2626)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3054..3074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3072..3369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3423..4217
                     /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(3477..3496)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4087..4106
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4869..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4906..4925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4960..4979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5008..5027)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5009..5027)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5041..5057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5041..5057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5054..5076)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5065..5081
                     /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(5089..5095,5096..5113,5114..5119))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5134..5155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5272..5289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5443..6031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5523..5542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6202..6993,6994..7062))
                     /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     6825..6844
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7063..7167)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7194..7622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7281..7300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7491..7512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctgg aagttctgtt ccaggggccc
     1021 cttgcggccg ccgatggcaa cgcggaaagc ctgtacatgc agggtatgga gctgtttgag
     1081 gaagcgctgc agaagtggga gcaagcgctg agcgttggtc aacgtggcga cagcggtagc
     1141 accccgatgc cgcgtgatgg tctgcgtaac ccggaaaccg cgagcgagcc gctgagcgag
     1201 ccggaaagcc agcgtaaaga gttcgcggaa aagctggaga gcctgctgca ccgtgcgtat
     1261 cacctgcaag aggaattcgg cagcaccttt ccggcggaca gcatgctgct ggatctggaa
     1321 cgtaccctga tgctgccgct gaccgagggt agcctgcgtc tgcgtgcgga cgatgaagac
     1381 agcctgacca gcgaggatag cttctttagc gcgaccgaac tgttcgagag cctgcagacc
     1441 ggcgactatc cgattccgct gagccgtccg gcggcggcgt atgaggaagc gctgcaactg
     1501 gtgaaagaag gccgtgttcc gtgccgtacc ctgcgtaccg agctgctggg ttgctacagc
     1561 gaccaggatt ttctggcgaa gctgcactgc gtgcgtcaag cgttcgaagg tctgctggag
     1621 gataaaagca accagctgtt ctttggcaaa gtgggtcgtc aaatggttac cggcctgatg
     1681 accaaggcgg agaagagccc gaagggtttt ctggagagct acgaggaaat gctgagctat
     1741 gcgctgcgtc cggaaacctg ggcgaccacc cgtctggagc tggaaggtcg tggcgtggtt
     1801 tgcatgagct tctttgacat cgttctggat tttattctga tggacgcgtt cgaagatctg
     1861 gagaacccgc cggcgagcgt gctggcggtt ctgcgtaacc gttggctgag cgacagcttc
     1921 aaagaaaccg cgctggcgac cgcgtgctgg agcgtgctga aggcgaaacg tcgtctgctg
     1981 atggttccgg atggtttcat cagccacttt tacagcgtga gcgaacacgt gagcccggtt
     2041 ctggcgttcg gctttctggg tccgaaaccg cagctggcgg aagtgtgcgc gttctttaag
     2101 caccagattg ttcaatacct gcgtgacatg ttcgacctgg ataacgttcg ttataccagc
     2161 ctgccggcgc tggcggatga tatcctgcag ctgagccgtc gtcgtagcga gattctgctg
     2221 ggttatctgg gtgtgccggc ggcgagcagc gcgggcgtta acggtgcgct gccgcgtgaa
     2281 aacggtccgc tgggcgagct gcaaggtggc ggtggtagtg gcggtggcgg cagcggaggt
     2341 ggcggtagtg ggggtggtgg aagcggcgga ggaggtagtc atcaccatca ccatcactaa
     2401 ggatcccggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     2461 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     2521 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     2581 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     2641 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     2701 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     2761 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     2821 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     2881 ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc
     2941 tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt
     3001 tgggtccact cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt
     3061 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3121 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3181 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3241 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3301 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3361 cttttttgga ggaattgatc agcttgggat ctgatcaaga gacaggatga ggatcgtttc
     3421 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3481 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3541 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3601 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3661 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3721 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3781 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3841 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3901 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3961 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4021 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4081 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4141 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4201 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4261 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4321 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4381 cgcccacccc gggctcgatc ccctcgcgag ttggttcagc tgctgcctga ggctggacga
     4441 cctcgcggag ttctaccggc agtgcaaatc cgtcggcatc caggaaacca gcagcggcta
     4501 tccgcgcatc catgcccccg aactgcagga gtggggaggc acgatggccg ctttggtcga
     4561 cccggacggg acgctcctgc gcctgataca gaacgaattg cttgcaggca tctcatgagt
     4621 gtgtcttccc gttttccgcc tgaggtcact gcgtggatgg agcgctggcg cctgctgcgc
     4681 gacggcgagc tgctcaccac ccactcgcca agctggaacc gtaaaaaggc cgcgttgctg
     4741 gcgtttttcc ataggctccg ccgatcataa tcagccatac cacatttgta gaggttttac
     4801 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4861 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4921 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4981 atgtatctta tcatgtctgg atcaattccc tatagtgagt cgtattaaat tcgtaatcat
     5041 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5101 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5161 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5221 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5281 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5341 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5401 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5461 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5521 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5581 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5641 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5701 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5761 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5821 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5881 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5941 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6001 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6061 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6121 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6181 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6241 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6301 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6361 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6421 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6481 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6541 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6601 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6661 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6721 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6781 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6841 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6901 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6961 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7021 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7081 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7141 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     7201 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     7261 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     7321 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     7381 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     7441 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     7501 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     7561 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     7621 ttttaacaaa atattaacgc ttacaattt
//
