LOCUS       sequence-75126-e        8902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human SDHB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8902)
  AUTHORS   Sancak Y, Markhard AL, Kitami T, Kovacs-Bogdan E, Kamer KJ, Udeshi 
            ND, Carr SA, Chaudhuri D, Clapham DE, Li AA, Calvo SE, Goldberger O,
            Mootha VK
  TITLE     EMRE is an Essential Component of the Mitochondrial Calcium 
            Uniporter Complex.
  JOURNAL   Science. 2013 Nov 14.
  PUBMED    24231807
REFERENCE   2  (bases 1 to 8902)
  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: "Science. 
            2013 Nov 14."
COMMENT     Sequence Label: pLYS5-SDHB-Flag
FEATURES             Location/Qualifiers
     source          1..8902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1239..1262
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1313..1430
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     1922..1941
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     LTR             3410..3643
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3715..3836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3752..3771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3806..3825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3876..4011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4031..4050)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4032..4050)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4060..4077)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4060..4076)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4069..4091)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4218..4673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4305..4324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4515..4536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4699..4803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4804..4872,4873..5664)
                     /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     complement(5022..5041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5835..6423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6324..6343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6577..6594
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6711..6732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6747..6752,6753..6770,6771..6777)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6785..6801
                     /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)."
     primer_bind     6790..6812
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6809..6825
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6809..6825
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6844..6864
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6846..6864
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6892..7118
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7119..7299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7346..7471
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7964..8197
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8382..8426
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     8724..8744
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcatggcggc ggtggtcgcc ctctccttga ggcgccggtt gccggccaca acccttggcg
      421 gagcctgcct gcaggcctcc cgaggagccc agacagctgc agccacagct ccccgtatca
      481 agaaatttgc catctatcga tgggacccag acaaggctgg agacaaacct catatgcaga
      541 cttatgaagt tgaccttaat aaatgtggcc ccatggtatt ggatgcttta atcaagatta
      601 agaatgaagt tgactctact ttgaccttcc gaagatcatg cagagaaggc atctgtggct
      661 cttgtgcaat gaacatcaat ggaggcaaca ctctagcttg cacccgaagg attgacacca
      721 acctcaataa ggtctcaaaa atctaccctc ttccacacat gtatgtgata aaggatcttg
      781 ttcccgattt gagcaacttc tatgcacagt acaaatccat tgagccttat ttgaagaaga
      841 aggatgaatc tcaggaaggc aagcagcagt atctgcagtc catagaagag cgtgagaaac
      901 tggacgggct ctacgagtgc attctctgtg cctgctgtag caccagctgc cccagctact
      961 ggtggaacgg agacaaatat ctggggcctg cagttcttat gcaggcctat cgctggatga
     1021 ttgactccag agatgacttc acagaggagc gcctggccaa gctgcaggac ccattctctc
     1081 tataccgctg ccacaccatc atgaactgca caaggacctg tcctaagggt ctgaatccag
     1141 ggaaagctat tgcagagatc aagaaaatga tggcaaccta taaggagaag aaagcttcag
     1201 ttcccggggg tggatctggt ggatctggtg gatctatgga ttacaaggat gacgatgaca
     1261 agtaaggtac cgaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa
     1321 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     1381 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     1441 acagggacag cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc
     1501 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
     1561 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
     1621 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
     1681 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
     1741 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     1801 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     1861 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     1921 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     1981 ggcctttcga cctgcatccc gccaccatga aaaagcctga actcaccgcg acgtctgtcg
     2041 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     2101 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     2161 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     2221 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     2281 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     2341 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     2401 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     2461 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     2521 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     2581 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     2641 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     2701 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     2761 ggcatccgga gcttgcagga tcgccgcggc tccggggcgt atatgctccg cattggtctt
     2821 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     2881 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     2941 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     3001 cgccccagca ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa caagagctga
     3061 tttcgagaac gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg
     3121 gccttacgct tggtggcaca gttctcgtcc acagttcgct aagctcgctc ggctgggtcg
     3181 cgggagggcc ggtcgcagtg attcaggccc ttctggattg tgttggtccc cagggcacga
     3241 ttgtcatgcc cacgcactcg ggtgatctga ctgatcccgc agattggaga tcgccgcccg
     3301 tgcctgccga ttgggtgcag atccgtcgac ctgcagccaa gctctttaag accaatgact
     3361 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     3421 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     3481 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     3541 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     3601 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     3661 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     3721 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3781 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3841 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3901 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3961 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     4021 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     4081 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4141 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4201 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4261 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4321 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4381 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4441 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4501 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4561 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4621 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4681 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4741 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4801 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4861 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4921 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4981 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5041 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5101 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5161 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5221 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5281 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5341 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5401 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5461 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5521 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5581 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5641 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5701 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5941 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6421 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6481 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6541 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6601 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6661 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6721 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6781 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     6841 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     6901 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     6961 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     7021 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     7081 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     7141 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7201 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7261 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     7321 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     7381 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     7441 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     7501 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     7561 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     7621 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     7681 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     7741 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7801 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     7861 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7921 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7981 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     8041 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     8101 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     8161 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     8221 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     8281 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     8341 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     8401 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     8461 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     8521 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     8581 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     8641 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     8701 ctcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
     8761 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     8821 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     8881 tatgttttaa aatggactat ca
//
