LOCUS       sequence-235949-        5398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GPPS and LimS, for the production of GPP and (S)-limonene,
            in Escherichia coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5398)
  AUTHORS   Jervis AJ, Carbonell P, Vinaixa M, Dunstan MS, Hollywood KA, 
            Robinson CJ, Rattray NJW, Yan C, Swainston N, Currin A, Sung R, 
            Toogood HS, Taylor S, Faulon JL, Breitling R, Takano E, Scrutton NS
  TITLE     Machine learning of designed translational control allows predictive
            pathway optimisation in Escherichia coli.
  JOURNAL   ACS Synth Biol. 2018 Dec 18. doi: 10.1021/acssynbio.8b00398.
  PUBMED    30563328
REFERENCE   2  (bases 1 to 5398)
  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: "ACS Synth 
            Biol. 2018 Dec 18. doi: 10.1021/acssynbio.8b00398."
COMMENT     Sequence Label: pGL
FEATURES             Location/Qualifiers
     source          1..5398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3336..3407
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3423..3450
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3608..4196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3688..3707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4284..4378)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(4408..5199,5200..5268))
                     /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     5031..5050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5269..5373)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcacgatctt
      721 aagttcaaga gggtaataat ggagttcgac ttcaacaaat acatggacag caaagccatg
      781 accgttaatg aagcactgaa taaagcaatt ccgctgcgtt atccgcagaa aatctatgaa
      841 agcatgcgtt atagcctgct ggcaggcggt aaacgtgttc gtccggttct gtgtattgca
      901 gcatgtgaac tggttggtgg caccgaagaa ctggcaattc cgaccgcatg tgcaattgaa
      961 atgattcata ccatgagcct gatgcatgat gatctgccgt gtattgataa tgatgacctg
     1021 cgtcgtggta aaccgaccaa tcataaaatc tttggtgaag ataccgcagt gaccgcaggt
     1081 aatgcactgc atagttatgc atttgaacat attgcagtga gcaccagcaa aaccgttggt
     1141 gcagatcgta ttctgcgtat ggttagcgaa ctgggtcgtg caaccggtag cgaaggtgtt
     1201 atgggtggtc agatggttga tattgcaagt gaaggtgatc cgagcattga tctgcagacc
     1261 ctggaatgga ttcatattca taaaaccgca atgctgctgg aatgtagcgt tgtttgtggt
     1321 gcaattattg gtggtgcaag cgaaattgtt attgaacgtg cccgtcgtta tgcacgttgt
     1381 gttggtctgc tgtttcaggt tgttgatgat attctggatg tgaccaaaag cagtgatgaa
     1441 ctgggcaaaa ccgcaggcaa agatctgatt agcgataaag caacctatcc gaaactgatg
     1501 ggtctggaaa aagccaaaga attttcagat gaactgctga atcgtgccaa aggtgaactg
     1561 agctgttttg atccggttaa agcagcaccg ctgctgggtc tggcagatta tgttgcattt
     1621 cgtcagaatt aagcttagag taaaactaag catagagaga gcgtactaat ggaacgtcgt
     1681 agcggtaact ataatccgag ccgttgggat gttaacttta ttcagagcct gctgagcgat
     1741 tacaaagagg ataaacacgt tattcgtgca agcgaactgg ttaccttagt taaaatggaa
     1801 ctggaaaaag aaaccgatca gattcgtcag ctggaactga ttgatgatct gcagcgtatg
     1861 ggtctgagcg atcattttca gaacgaattt aaagaaatcc tgagcagcat ctatttagat
     1921 caccactatt acaaaaaccc gtttccgaaa gaagaacgcg atctgtatag caccagcctg
     1981 gcatttcgtc tgctgcgtga acatggtttt caggttgcac aagaagtttt cgacagcttc
     2041 aaaaatgaag agggcgagtt taaagaatcc ctgtcagatg atacccgtgg tctgctgcag
     2101 ctgtatgaag ccagctttct gctgaccgaa ggtgaaacca ccctggaaag cgcacgtgaa
     2161 tttgcaacca aatttctgga agaaaaagtg aacgaaggtg gtgttgacgg cgatctgctg
     2221 acacgtattg catatagcct ggatattccg ctgcattggc gtattaaacg tccgaatgca
     2281 ccggtttgga ttgaatggta tcgcaaacgt ccggatatga atccggttgt tctggaactg
     2341 gcaattctgg atctgaatat tgttcaggca cagtttcaag aggaactgaa agaaagtttt
     2401 cgttggtggc gtaataccgg ctttgttgaa aaactgccgt ttgcacgtga tcgtctggtt
     2461 gaatgctatt tttggaacac cggtattatt gaaccgcgtc agcatgccag cgcacgtatt
     2521 atgatgggta aagttaatgc actgatcacc gtgatcgatg atatctatga cgtttatggt
     2581 acactggaag aactggaaca gtttacagat ctgattcgtc gttgggatat caacagcatt
     2641 gatcagctgc cggattatat gcagctgtgt tttctggcac tgaacaactt tgttgatgat
     2701 accagctatg atgtgatgaa agaaaaaggc gttaacgtga ttccgtatct gcgtcagagc
     2761 tgggttgatc tggcagataa atacatggtt gaagcccgtt ggttttatgg tggtcataaa
     2821 ccgagcctgg aagaatatct ggaaaatagc tggcagagca ttagcggtcc gtgtatgctg
     2881 acccatatct tttttcgtgt taccgatagc ttcaccaaag aaacagttga tagcctgtat
     2941 aaataccacg atctggttcg ttggagcagc tttgttctgc gtctggccga tgatctgggc
     3001 accagcgttg aagaggttag ccgcggcgat gttccgaaaa gcctgcagtg ttatatgagt
     3061 gattataatg ccagcgaagc cgaagcacgt aaacatgtta aatggctgat tgccgaagtg
     3121 tggaaaaaaa tgaacgcaga acgtgttagc aaagatagcc cgtttggcaa agattttatt
     3181 ggttgtgcag tggatctggg tcgtatggca cagctgatgt atcataatgg tgatggtcat
     3241 ggcacccagc atccgattat tcatcagcag atgacccgta ccctgtttga accgtttgca
     3301 taaggatcca aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga
     3361 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     3421 ctggctcacc ttcgggtggg cctttctgcg tttataccta gggcgttcgg ctgcggcgag
     3481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3781 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4021 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4261 ttttggtcat gactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     4321 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     4381 agcgagctcg taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4441 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4501 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4561 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4621 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4681 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4741 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4801 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4861 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4921 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4981 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5041 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5101 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5161 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5221 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5341 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacct
//
