LOCUS       sequence-298692-        6996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of biosynthetic enzyme STS in fusion with CC segment P4S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6996)
  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 6996)
  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:P4S
FEATURES             Location/Qualifiers
     source          1..6996
                     /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(1618..1632)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             complement(1648..1677)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1690..1712
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1727..1751
                     /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(1751..1770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1752..1770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1854..1873)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2079..2156
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2157..3239
                     /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(2176..2195)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4551..4742
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4736..4758)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4844..4983
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4899..4918
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4998..5015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5169..5757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5249..5268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5928..6719,6720..6788))
                     /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     6551..6570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6789..6893)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6961..6979
                     /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 atccttagcc gtatttaata ctctgcaact tttgcttgat
      361 ttgttgtaat ttctgttcat tttgttgcag ttcctcttcg ttggattgaa gctcgtcctc
      421 tggcgagctg ccgctgccat tggtaaccat cggaatgcta tgcagaacaa cggtttcaat
      481 ggtcagaccc ggaccaaaac caaacagaac accccaatcc agaccttcac cggtggttgc
      541 acgttcacct ttcaggcttt ttttgcgcat ttcatccaga ataaacagaa cacatgcgct
      601 gctcatatta ccatattcgc tcagaacatg acgggttgct tccagtttct ttttgtccag
      661 gttcagtttt gcttcaactg catccagaat tgccggacca cccggatgtg caatccaaaa
      721 caggctattc caatcgctaa tacccagcgg atcaaatgcc tgggtcagac atttttcaac
      781 attttcgcta atcagggtcg gaacattcgg ccacagatga aaggtcagac caacttcacg
      841 cagattacct gcaattgcac cggcactatt cggaataaag gtctgtgctg cgctaaccag
      901 ctgaaacagc ggacgttcaa tgctaatatc cggatcgcta ccaacaataa ctgctgcgct
      961 accatcacca aacagtgcct gaccaaccag gctatccagt gcatcttcgc tcggaccacg
     1021 aaaggtaaca acggtaattt cgctacaaac aaccagaaca cgtgcaccgg cattattttc
     1081 tgccagatct tttgcggtac gcagaacggt gccaccggca taacaaccct gatgatacag
     1141 cataacacga cgaacgctcg gttccagacc cagcagattt gccagtttat aatctgcacc
     1201 aggcatttca acaccactgg tggtacaaaa aaccagatgg gtaattttgc ttttcggctg
     1261 accccattct ttcagggctt tcagtgctgc ttctttaccc agtttcggaa cttctgcggt
     1321 gataatttct tgacgaatat tcaggctcgg tgccatatat gcaccaatat tcggatgttc
     1381 ttccagcatt tcttcggtca ggtggatata gcgttttttg atcatgcttt tatcgcagat
     1441 gcggttaaat ttctttttca gtgcggtcat gtgttcgctt ttggtcactt taaagtaata
     1501 atcggcataa tcgctctgat aaacacaatg atccggtgtt gcggtgccaa ttgccagaat
     1561 ggttgccgga ccttttgcac gctgtgcatt acgaaattct tcaacgcttg ccatatgctt
     1621 gtcgtcgtcg tcgatatggc cgctgctgtg atgatgatga tgatgatgat gatgatggcc
     1681 catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg
     1741 ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta
     1801 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     1861 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     1921 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     1981 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     2041 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
     2101 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     2161 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     2221 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     2281 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     2341 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     2401 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     2461 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     2521 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     2581 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     2641 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     2701 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     2761 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     2821 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     2881 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     2941 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     3001 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     3061 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     3121 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     3181 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3241 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
     3301 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     3361 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     3421 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     3481 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     3541 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     3601 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     3661 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     3721 ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg
     3781 atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta
     3841 ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
     3901 acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
     3961 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     4021 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct
     4081 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     4141 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     4201 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     4261 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat
     4321 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct
     4381 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
     4441 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
     4501 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
     4561 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     4621 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     4681 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     4741 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4801 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     4861 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     4921 gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4981 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5041 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5101 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5341 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5581 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5641 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5881 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5941 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6001 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6061 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6121 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6181 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6241 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6301 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6361 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6421 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6481 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6541 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6601 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6661 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6721 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6781 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6901 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6961 aataggcgta tcacgaggcc ctttcgtctt caagaa
//
