LOCUS       sequence-308955-        6990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses biosynthetic enzyme STS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  AUTHORS   Fink T, Stevovic B, Verwaal R, Roubos JA, Gaber R, Bencina M, Jerala
            R, Gradisar H
  TITLE     Metabolic enzyme clustering by coiled coils improves the 
            biosynthesis of resveratrol and mevalonate.
  JOURNAL   AMB Express. 2020 May 24;10(1):97. doi: 10.1186/s13568-020-01031-5.
  PUBMED    32448937
REFERENCE   2  (bases 1 to 6990)
  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: "AMB 
            Express."; date: "2020-05-24"; volume: "10(1):97. doi"; pages: " 
            10.1186/s13568-020-01031-5"
COMMENT     Sequence Label: pET19b_STS
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1612..1626)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             complement(1642..1671)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1684..1706
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1721..1745
                     /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     complement(1745..1764)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1746..1764)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1848..1867)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2073..2150
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2151..3233
                     /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     complement(2170..2189)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4545..4736
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4730..4752)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4838..4977
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4893..4912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4992..5009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5163..5751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5243..5262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5922..6713,6714..6782))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6545..6564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6783..6887)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6955..6973
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atccttaacg tttcaaatac tcaatcttgc gctctaattc
      361 gcggttttcc tcctccaaac gctcgatttc gcgctttaac tcctcattct tgtcctctgg
      421 ggagccgctg ccattggtaa ccatcggaat gctatgcaga acaacggttt caatggtcag
      481 acccggacca aaaccaaaca gaacacccca atccagacct tcaccggtgg ttgcacgttc
      541 acctttcagg ctttttttgc gcatttcatc cagaataaac agaacacatg cgctgctcat
      601 attaccatat tcgctcagaa catgacgggt tgcttccagt ttctttttgt ccaggttcag
      661 ttttgcttca actgcatcca gaattgccgg accacccgga tgtgcaatcc aaaacaggct
      721 attccaatcg ctaataccca gcggatcaaa tgcctgggtc agacattttt caacattttc
      781 gctaatcagg gtcggaacat tcggccacag atgaaaggtc agaccaactt cacgcagatt
      841 acctgcaatt gcaccggcac tattcggaat aaaggtctgt gctgcgctaa ccagctgaaa
      901 cagcggacgt tcaatgctaa tatccggatc gctaccaaca ataactgctg cgctaccatc
      961 accaaacagt gcctgaccaa ccaggctatc cagtgcatct tcgctcggac cacgaaaggt
     1021 aacaacggta atttcgctac aaacaaccag aacacgtgca ccggcattat tttctgccag
     1081 atcttttgcg gtacgcagaa cggtgccacc ggcataacaa ccctgatgat acagcataac
     1141 acgacgaacg ctcggttcca gacccagcag atttgccagt ttataatctg caccaggcat
     1201 ttcaacacca ctggtggtac aaaaaaccag atgggtaatt ttgcttttcg gctgacccca
     1261 ttctttcagg gctttcagtg ctgcttcttt acccagtttc ggaacttctg cggtgataat
     1321 ttcttgacga atattcaggc tcggtgccat atatgcacca atattcggat gttcttccag
     1381 catttcttcg gtcaggtgga tatagcgttt tttgatcatg cttttatcgc agatgcggtt
     1441 aaatttcttt ttcagtgcgg tcatgtgttc gcttttggtc actttaaagt aataatcggc
     1501 ataatcgctc tgataaacac aatgatccgg tgttgcggtg ccaattgcca gaatggttgc
     1561 cggacctttt gcacgctgtg cattacgaaa ttcttcaacg cttgccatat gcttgtcgtc
     1621 gtcgtcgata tggccgctgc tgtgatgatg atgatgatga tgatgatgat ggcccatggt
     1681 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
     1741 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
     1801 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     1861 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     1921 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     1981 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     2041 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
     2101 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     2161 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     2221 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     2281 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     2341 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     2401 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     2461 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     2521 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     2581 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2641 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2701 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2761 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2821 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     2881 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     2941 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     3001 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     3061 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     3121 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3241 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     3301 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     3361 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     3421 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     3481 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     3541 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     3601 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     3661 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     3721 cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc gctgatcgtc
     3781 acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc
     3841 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     3901 acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga attggagcca
     3961 atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg
     4021 cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg tcctggccac
     4081 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     4141 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     4201 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     4261 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     4321 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     4381 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     4441 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     4501 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
     4561 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
     4621 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
     4681 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4741 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4801 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     4861 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     4921 gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     4981 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5581 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     6241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     6541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6901 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6961 cgtatcacga ggccctttcg tcttcaagaa
//
