LOCUS       sequence-386297-        6092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of myc-tagged human Nix with an aspartic acid 
            substitution of serine-212. Based on Myc-Nix (#100795).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6092)
  AUTHORS   da Silva Rosa SC, Martens MD, Field JT, Nguyen L, Kereliuk SM, Hai 
            Y, Chapman D, Diehl-Jones W, Aliani M, West AR, Thliveris J, Ghavami
            S, Rampitsch C, Dolinsky VW, Gordon JW
  TITLE     BNIP3L/Nix-induced mitochondrial fission, mitophagy, and impaired 
            myocyte glucose uptake are abrogated by PRKA/PKA phosphorylation.
  JOURNAL   Autophagy. 2021 Sep;17(9):2257-2272. doi: 
            10.1080/15548627.2020.1821548. Epub 2020 Oct 12.
  PUBMED    33044904
REFERENCE   2  (bases 1 to 6092)
  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"; doi: 
            "10.1080/15548627.2020.1821548"; journalName: "Autophagy"; date: 
            "2021-09"; volume: "17"; issue: "9"; pages: "2257-2272"
COMMENT     Sequence Label: Myc-Nix-S212D
FEATURES             Location/Qualifiers
     source          1..6092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..933
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1645..1663)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1646..1663)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1683..1700)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1689..1913
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1959..2387
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2046..2065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2256..2277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2396..2416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2401..2730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2581..2716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2643..2662
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2797..3591
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVTCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2851..2870)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3461..3480
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3765..3886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3802..3821)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3856..3875
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3935..3951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3935..3951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3948..3970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3959..3975
                     /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(3983..3989,3990..4007,4008..4013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4028..4049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4166..4183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4337..4925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4417..4436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5096..5887,5888..5956))
                     /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     5719..5738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5957..6061)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 atggagcaga agctgatctc cgaggaggac ctgggatcca tcgatgtcga cgaattcatg
      961 tcgtcccacc tagtcgagcc gccgccgccc ctgcacaaca acaacaacaa ctgcgaggaa
     1021 aatgagcagt ctctgccccc gccggccggc ctcaacagtt cctgggtgga gctacccatg
     1081 aacagcagca atggcaatga taatggcaat gggaaaaatg gggggctgga acacgtacca
     1141 tcctcatcct ccatccacaa tggagacatg gagaagattc ttttggatgc acaacatgaa
     1201 tcaggacaga gtagttccag aggcagttct cactgtgaca gcccttcgcc acaagaagat
     1261 gggcagatca tgtttgatgt ggaaatgcac accagcaggg accatagctc tcagtcagaa
     1321 gaagaagttg tagaaggaga gaaggaagtc gaggctttga agaaaagtgc ggactgggta
     1381 tcagactggt ccagtagacc cgaaaacatt ccacccaagg agttccactt cagacaccct
     1441 aaacgttctg tgtctttaag catgaggaaa agtggagcca tgaagaaagg gggtattttc
     1501 tccgcagaat ttctgaaggt gttcattcca tctctcttcc tttctcatgt tttggctttg
     1561 gggctaggca tctatattgg aaagcgactg gacacaccct ctgccagcac ctactgactc
     1621 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     1681 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1741 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1801 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1861 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     1921 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     1981 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2041 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2101 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2161 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2221 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2281 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2341 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2401 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2461 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2521 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2581 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2641 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     2701 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     2761 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     2821 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     2881 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     2941 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3001 tggctggcca cgacgggcgt tacttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3061 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3121 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3181 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3241 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3301 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3361 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3421 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3481 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3541 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3601 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3661 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3721 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     3781 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3841 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     3901 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3961 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4021 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4081 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4141 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4741 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5401 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5701 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6061 gcacatttcc ccgaaaagtg ccacctgacg tc
//
