LOCUS       sequence-220630-        5291 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 5291)
  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 5291)
  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..5291
                     /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        2086..2177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2178..2246,2247..3038)
                     /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(2396..2415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3209..3797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3698..3717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3951..3968
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3983..4123
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4049..4068)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4209..4231
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4387..5265)
                     /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 actcttttgt ttatttttct
     2101 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2161 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2221 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2281 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2341 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2401 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2461 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2521 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2581 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2641 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2701 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2761 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2821 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2881 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2941 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3001 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3061 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3121 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3181 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3241 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3301 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3361 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3421 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3481 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3541 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3601 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3661 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3721 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3781 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3841 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3901 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3961 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4021 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4081 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     4141 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4201 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4261 gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga
     4321 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt
     4381 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
     4441 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
     4501 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
     4561 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
     4621 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
     4681 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
     4741 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
     4801 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
     4861 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
     4921 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
     4981 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
     5041 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
     5101 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
     5161 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
     5221 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
     5281 gccattcaga g
//
