LOCUS       sequence-193039-        5990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nix dominant negative.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  AUTHORS   Mughal W, Martens M, Field J, Chapman D, Huang J, Rattan S, Hai Y, 
            Cheung KG, Kereliuk S, West AR, Cole LK, Hatch GM, Diehl-Jones W, 
            Keijzer R, Dolinsky VW, Dixon IM, Parmacek MS, Gordon JW
  TITLE     Myocardin regulates mitochondrial calcium homeostasis and prevents 
            permeability transition.
  JOURNAL   Cell Death Differ. 2018 Mar 6. pii: 10.1038/s41418-018-0073-z. doi: 
            10.1038/s41418-018-0073-z.
  PUBMED    29511336
REFERENCE   2  (bases 1 to 5990)
  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: "Cell Death 
            Differ. 2018 Mar 6. pii: 10.1038/s41418-018-0073-z. doi: 
            10.1038/s41418-018-0073-z."
COMMENT     Sequence Label: Myc-Nix-delta TM
FEATURES             Location/Qualifiers
     source          1..5990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        21..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     649..668
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        649..667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             690..719
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1329..1347)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1330..1347)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1367..1384)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1373..1597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1643..2071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1730..1749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1940..1961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2080..2100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2085..2414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2265..2400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2327..2346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2481..3275
                     /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(2535..2554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3145..3164
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3449..3570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3486..3505)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3540..3559
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3619..3635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3619..3635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3632..3654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3643..3659
                     /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(3667..3673,3674..3691,3692..3697))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3712..3733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3850..3867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4021..4609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4101..4120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4780..5571,5572..5640))
                     /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     5403..5422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5641..5745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5820..5839)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      661 ctatagggag acccaagctt ggtaccatgg agcagaagct gatctccgag gaggacctgg
      721 gatccatcga tgtcgacgaa ttcatgtcgt cccacctagt cgagccgccg ccgcccctgc
      781 acaacaacaa caacaactgc gaggaaaatg agcagtctct gcccccgccg gccggcctca
      841 acagttcctg ggtggagcta cccatgaaca gcagcaatgg caatgataat ggcaatggga
      901 aaaatggggg gctggaacac gtaccatcct catcctccat ccacaatgga gacatggaga
      961 agattctttt ggatgcacaa catgaatcag gacagagtag ttccagaggc agttctcact
     1021 gtgacagccc ttcgccacaa gaagatgggc agatcatgtt tgatgtggaa atgcacacca
     1081 gcagggacca tagctctcag tcagaagaag aagttgtaga aggagagaag gaagtcgagg
     1141 ctttgaagaa aagtgcggac tgggtatcag actggtccag tagacccgaa aacattccac
     1201 ccaaggagtt ccacttcaga caccctaaac gttctgtgtc tttaagcatg aggaaaagtg
     1261 gagccatgaa gaaagggggt attttctccg cagaattttg actcgagcat gcatctagag
     1321 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     1381 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1441 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1501 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1561 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     1621 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1681 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1741 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1801 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1861 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1921 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1981 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2041 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2101 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2161 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2221 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2281 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2341 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2401 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     2461 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     2521 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     2581 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     2641 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     2701 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     2761 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     2821 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     2881 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     2941 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3001 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3061 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3121 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3181 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3241 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3301 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3361 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3421 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     3481 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3541 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     3601 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4441 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5761 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     5821 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     5881 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     5941 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
//
