LOCUS       Exported                7233 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Hetero expression of His-tagged human SDHA.
ACCESSION   .
VERSION     .
KEYWORDS    H6hSDHA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7233)
  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 7233)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7233
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2084..2102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2103..2127
                     /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             2179..2223
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2261..2279)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2275..2322
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2359..2814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2446..2465)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2656..2677
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2929..3789)
                     /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     3552..3571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3790..3882)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3963..4551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4452..4471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4705..4722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4737..4879
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4805..4824)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4965..4987
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4981..5172)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5744..6826)
                     /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     6788..6807
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6827..6904)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7110..7129
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7217..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cggcatctgc
      121 taaagtttca gattccattt ctgctcagta tccagtagtg gatcatgaat ttgatgcagt
      181 ggtggtaggc gctggagggg caggcttgcg agctgcattt ggcctttctg aggcagggtt
      241 taatacagca tgtgttacca agctgtttcc taccaggtca cacactgttg cagcacaggg
      301 aggaatcaat gctgctctgg ggaacatgga ggaggacaac tggaggtggc atttctacga
      361 caccgtgaag ggctccgact ggctggggga ccaggatgcc atccactaca tgacggagca
      421 ggcccccgcc gccgtggtcg agctagaaaa ttatggcatg ccgtttagca gaactgaaga
      481 tgggaagatt tatcagcgtg catttggtgg acagagcctc aagtttggaa agggcgggca
      541 ggcccatcgg tgctgctgtg tggctgatcg gactggccac tcgctattgc acaccttata
      601 tggaaggtct ctgcgatatg ataccagcta ttttgtggag tattttgcct tggatctcct
      661 gatggagaat ggggagtgcc gtggtgtcat cgcactgtgc atagaggacg ggtccatcca
      721 tcgcataaga gcaaagaaca ctgttgttgc cacaggaggc tacgggcgca cctacttcag
      781 ctgcacgtct gcccacacca gcactggcga cggcacggcc atgatcacca gggcaggcct
      841 tccttgccag gacctagagt ttgttcagtt ccaccccaca ggcatatatg gtgctggttg
      901 tctcattacg gaaggatgtc gtggagaggg aggcattctc attaacagtc aaggcgaaag
      961 gtttatggag cgatacgccc ctgtcgcgaa ggacctggcg tctagagatg tggtgtctcg
     1021 gtccatgact ctggagatcc gagaaggaag aggctgtggc cctgagaaag atcacgtcta
     1081 cctgcagctg caccacctac ctccagagca gctggccacg cgcctgcctg gcatttcaga
     1141 gacagccatg atcttcgctg gcgtggacgt cacgaaggag ccgatccctg tcctccccac
     1201 cgtgcattat aacatgggcg gcattcccac caactacaag gggcaggtcc tgaggcacgt
     1261 gaatggccag gatcagattg tgcccggcct gtacgcctgt ggggaggccg cctgtgcctc
     1321 ggtacatggt gccaaccgcc tcggggcaaa ctcgctcttg gacctggttg tctttggtcg
     1381 ggcatgtgcc ctgagcatcg aagagtcatg caggcctgga gataaagtcc ctccaattaa
     1441 accaaacgct ggggaagaat ctgtcatgaa tcttgacaaa ttgagatttg ctgatggaag
     1501 cataagaaca tcggaactgc gactcagcat gcagaagtca atgcaaaatc atgctgccgt
     1561 gttccgtgtg ggaagcgtgt tgcaagaagg ttgtgggaaa atcagcaagc tctatggaga
     1621 cctaaagcac ctgaagacgt tcgaccgggg aatggtctgg aacacggacc tggtggagac
     1681 cctggagctg cagaacctga tgctgtgtgc gctgcagacc atctacggag cagaggcacg
     1741 gaaggagtca cggggcgcgc atgccaggga agactacaag gtgcggattg atgagtacga
     1801 ttactccaag cccatccagg ggcaacagaa gaagcccttt gaggagcact ggaggaagca
     1861 caccctgtcc tatgtggacg ttggcactgg gaaggtcact ctggaatata gacccgtgat
     1921 cgacaaaact ttgaacgagg ctgactgtgc caccgtcccg ccagccattc gctcctactg
     1981 agaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc
     2041 gaacagaaag taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata
     2101 ggggaattgt gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga
     2161 gatatactcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac
     2221 tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca
     2281 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     2341 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2401 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2461 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2521 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2581 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2641 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2701 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2761 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttctggcg
     2821 gcacgatggc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3541 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3781 aatactcata ctcttccttt ttcaatcatg attgaagcat ttatcagggt tattgtctca
     3841 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggtcat gaccaaaatc
     3901 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3961 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4021 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4081 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4141 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4201 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4261 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4321 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4381 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4441 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4501 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4561 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4621 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4681 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4741 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc
     4801 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     4861 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4921 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4981 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     5041 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     5101 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     5161 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     5221 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     5281 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     5341 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     5401 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     5461 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     5521 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     5581 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc
     5641 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     5701 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     5761 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5821 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     5881 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     5941 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     6001 tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg
     6061 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     6121 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     6181 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     6241 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     6301 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     6361 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     6421 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     6481 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     6541 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     6601 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     6661 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     6721 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     6781 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     6841 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     6901 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     6961 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     7021 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     7081 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     7141 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat
     7201 ctcgatcccg cgaaattaat acgactcact ata
//
