LOCUS       sequence-217502-        7230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hetero expression of His-tagged human SDHA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7230)
  AUTHORS   Zafreen L, Walker-Kopp N, Huang LS, Berry E
  TITLE     In-vitro, SDH5-dependent flavinylation of immobilized human 
            respiratory complex II flavoprotein.
  JOURNAL   Arch Biochem Biophys. 2016 Aug 15;604:47-56. doi: 
            10.1016/j.abb.2016.06.006. Epub 2016 Jun 11.
  PUBMED    27296776
REFERENCE   2  (bases 1 to 7230)
  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: "Arch 
            Biochem Biophys."; date: "2016-08-15"; volume: "604:47-56. doi"; 
            pages: " 10.1016/j.abb.2016.06.006. Epub 2016 Jun 11"
COMMENT     Sequence Label: H6hSDHA
FEATURES             Location/Qualifiers
     source          1..7230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1779..2861)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2823..2842
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2862..2939)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3145..3164
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3252..3270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3271..3295
                     /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)."
     RBS             3310..3332
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3351..3368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        5352..5370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5371..5395
                     /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)."
     CDS             5447..5491
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(5529..5547)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5543..5590
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5627..6082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5714..5733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5924..5945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(6197..6988,6989..7057))
                     /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     6820..6839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7058..7150)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     1321 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     1381 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     1441 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     1501 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     1561 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     1621 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgctagtc atgccccgcg
     1681 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     1741 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1861 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     1921 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     1981 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     2041 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     2101 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     2161 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     2221 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     2281 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     2341 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     2401 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     2461 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     2521 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     2581 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     2641 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     2701 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     2761 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     2821 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     2881 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     2941 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     3001 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     3061 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     3121 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     3181 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga
     3241 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     3301 agaaataatt ttgtttaact ttaagaagga gatataccat gggcagcagc catcaccatc
     3361 atcaccacag ccaggatccg gcatctgcta aagtttcaga ttccatttct gctcagtatc
     3421 cagtagtgga tcatgaattt gatgcagtgg tggtaggcgc tggaggggca ggcttgcgag
     3481 ctgcatttgg cctttctgag gcagggttta atacagcatg tgttaccaag ctgtttccta
     3541 ccaggtcaca cactgttgca gcacagggag gaatcaatgc tgctctgggg aacatggagg
     3601 aggacaactg gaggtggcat ttctacgaca ccgtgaaggg ctccgactgg ctgggggacc
     3661 aggatgccat ccactacatg acggagcagg cccccgccgc cgtggtcgag ctagaaaatt
     3721 atggcatgcc gtttagcaga actgaagatg ggaagattta tcagcgtgca tttggtggac
     3781 agagcctcaa gtttggaaag ggcgggcagg cccatcggtg ctgctgtgtg gctgatcgga
     3841 ctggccactc gctattgcac accttatatg gaaggtctct gcgatatgat accagctatt
     3901 ttgtggagta ttttgccttg gatctcctga tggagaatgg ggagtgccgt ggtgtcatcg
     3961 cactgtgcat agaggacggg tccatccatc gcataagagc aaagaacact gttgttgcca
     4021 caggaggcta cgggcgcacc tacttcagct gcacgtctgc ccacaccagc actggcgacg
     4081 gcacggccat gatcaccagg gcaggccttc cttgccagga cctagagttt gttcagttcc
     4141 accccacagg catatatggt gctggttgtc tcattacgga aggatgtcgt ggagagggag
     4201 gcattctcat taacagtcaa ggcgaaaggt ttatggagcg atacgcccct gtcgcgaagg
     4261 acctggcgtc tagagatgtg gtgtctcggt ccatgactct ggagatccga gaaggaagag
     4321 gctgtggccc tgagaaagat cacgtctacc tgcagctgca ccacctacct ccagagcagc
     4381 tggccacgcg cctgcctggc atttcagaga cagccatgat cttcgctggc gtggacgtca
     4441 cgaaggagcc gatccctgtc ctccccaccg tgcattataa catgggcggc attcccacca
     4501 actacaaggg gcaggtcctg aggcacgtga atggccagga tcagattgtg cccggcctgt
     4561 acgcctgtgg ggaggccgcc tgtgcctcgg tacatggtgc caaccgcctc ggggcaaact
     4621 cgctcttgga cctggttgtc tttggtcggg catgtgccct gagcatcgaa gagtcatgca
     4681 ggcctggaga taaagtccct ccaattaaac caaacgctgg ggaagaatct gtcatgaatc
     4741 ttgacaaatt gagatttgct gatggaagca taagaacatc ggaactgcga ctcagcatgc
     4801 agaagtcaat gcaaaatcat gctgccgtgt tccgtgtggg aagcgtgttg caagaaggtt
     4861 gtgggaaaat cagcaagctc tatggagacc taaagcacct gaagacgttc gaccggggaa
     4921 tggtctggaa cacggacctg gtggagaccc tggagctgca gaacctgatg ctgtgtgcgc
     4981 tgcagaccat ctacggagca gaggcacgga aggagtcacg gggcgcgcat gccagggaag
     5041 actacaaggt gcggattgat gagtacgatt actccaagcc catccagggg caacagaaga
     5101 agccctttga ggagcactgg aggaagcaca ccctgtccta tgtggacgtt ggcactggga
     5161 aggtcactct ggaatataga cccgtgatcg acaaaacttt gaacgaggct gactgtgcca
     5221 ccgtcccgcc agccattcgc tcctactgag aattcgagct cggcgcgcct gcaggtcgac
     5281 aagcttgcgg ccgcataatg cttaagtcga acagaaagta atcgtattgt acacggccgc
     5341 ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc
     5401 ttagtatatt agttaagtat aagaaggaga tatactcgag tctggtaaag aaaccgctgc
     5461 tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc
     5521 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     5581 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     5641 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5701 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5761 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5821 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5881 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5941 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6001 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     6061 aaaatattaa cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct
     6121 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6181 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6241 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6301 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6361 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6421 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6481 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6541 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6601 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6661 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6721 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6781 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6841 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6901 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6961 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7021 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat
     7081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7141 aataaacaaa taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7201 agaccccgta gaaaagatca aaggatcttc
//
