LOCUS       sequence-205434-        5292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heterologous expression of HMGR of S. solfataricus in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5292)
  AUTHORS   Schulz F, Dirkmann M, Nowack J
  TITLE     An in vitro Biosynthesis of Sesquiterpenes starting from Acetic 
            Acid.
  JOURNAL   Chembiochem. 2018 Aug 7. doi: 10.1002/cbic.201800128.
  PUBMED    30085399
REFERENCE   2  (bases 1 to 5292)
  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: 
            "Chembiochem. 2018 Aug 7. doi: 10.1002/cbic.201800128."
COMMENT     Sequence Label: pBADM11-HMGR
FEATURES             Location/Qualifiers
     source          1..5292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             325..342
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             373..393
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(534..545)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1692..1709)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1862..1948
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2040..2067
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2087..2178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2179..2247,2248..3039)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2397..2416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3210..3798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3699..3718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3952..3969
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3984..4124
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4050..4069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4210..4232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4388..5266)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gaaacatcac catcaccatc accccatgag cgattacgac
      361 atccccacta ctgagaatct ttattttcag ggcgccatgg cgatgaaaat tgatgaagtg
      421 gttgaaaagt tagtaaaggg agaaatttct tttcatgaag ttgacaattt attagaagca
      481 aatgctgcaa tggttgcgag gaggcttgca ttagagaaga ttgtaggagt cggtcttccc
      541 tcaattggtt ctacagttat tgattatagt gagataaaaa ataaaaatgc cgaaaatgtg
      601 attggagcaa tacaaatacc attaggtata gtggggccaa ttagagtcaa tggtgattac
      661 gcaaagggcg atttctatgt acctatggcg acgacagaag gagcgctaat agccagtgta
      721 aataggggta tcaaggctgt tacactaagt gggggagtta gggctaaggt tttaaaggat
      781 gaaatgacga gagcccccgt ttttaagttt gattcaatag aacagatacc gaattttcta
      841 aagttcatag aggagaatct tgagaagata aggaatatag ccaattctac ctctcatcat
      901 ggtaaattaa aatccattac acctttcgta ctaggcaata atgtttggct tagattttcc
      961 tttgaaactg gtgatgctat ggggatgaac atggttacga tagcagttga aaaagtatgt
     1021 gagtttatag aggaaaactt cccatcagct gattgtttag ctgtaagcgg aaatatgtgt
     1081 agtgataaga aacagaccaa tgtcaatagc ctcttcggta gaggtaaaac tgtagtggct
     1141 gaggctctca tcaagaagga tgttataagg aatatattac actctaatgc tcaactaatt
     1201 catgatataa atctacggaa gaattggctc gggacagcaa gagcaggatc cttatctcaa
     1261 tttaatgctc atttcgcaaa catagtcaca gcaatcttta tagctactgg ccaagatgta
     1321 gctcaaatag ttgagagtag ttctggatat acatggactg aagtaagagg agaggactta
     1381 tatatatcag ttacactgcc ctcgttagaa gtaggaactg taggaggagg tacaaggtta
     1441 cctacccaga aagaagccct atccattatg ggagtttacg gcagtggaaa tccacctggc
     1501 tctaatgcta agaaattagc tgagatcata gcttctactg tactatctgg ggaacttaat
     1561 ctcctagctg ctttatccaa caaggagtta ggtaaggctc acgctaaact tggaagagct
     1621 atgaaagtct aataagcttg cggccgcact cgagagcttg gctgttttgg cggatgagag
     1681 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     1741 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     1801 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     1861 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1921 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     1981 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     2041 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttttg tttatttttc
     2101 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2161 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2221 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2281 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2341 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2401 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2461 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2521 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2581 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2641 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2701 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2761 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2821 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2881 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2941 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3001 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3061 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3121 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3181 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3241 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3301 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3361 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3421 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3481 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3541 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3601 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3661 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3721 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3781 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3841 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3901 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3961 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4021 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4081 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     4141 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4201 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4261 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg
     4321 acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg
     4381 ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     4441 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     4501 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     4561 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     4621 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     4681 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     4741 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     4801 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     4861 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     4921 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     4981 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     5041 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     5101 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     5161 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     5221 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     5281 cgccattcag ag
//
