LOCUS       pTrc-trGPPS(CO)-LS                6704 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pTrc-trGPPS(CO)-LS: A plasmid that has truncated codon optimized 
            geranylpyrophosphate synthase and truncated limonene synthase.
ACCESSION   .
VERSION     .
KEYWORDS    pTrc-trGPPS(CO)-LS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  AUTHORS   Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
            CJ, Lee TS
  TITLE     Metabolic engineering of Escherichia coli for limonene and perillyl 
            alcohol production.
  JOURNAL   Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
  PUBMED    23727191
REFERENCE   2  (bases 1 to 6704)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..6704
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        196..225
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    233..249
                     /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     238..260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(2890..2907)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3060..3146
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3238..3265
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3285..3376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3377..4237
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3595..3614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4408..4996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4897..4916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5150..5167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5182..5322
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5248..5267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5408..5430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5508..5585
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5586..6668
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5605..5624)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctggtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
       61 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
      121 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
      181 ttctgaaatg agctgttgac aattaatcat ccggctcgta taatgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagacca tggaatttga cttcaacaaa tacatggact
      301 ccaaagcgat gacggtaaat gaagcactga acaaagcgat ccctctgcgt tatccgcaga
      361 aaatctacga aagcatgcgt tacagcctgc tggcaggcgg caagcgtgtt cgtccggttc
      421 tgtgtattgc cgcatgtgaa ctggtaggtg gtaccgaaga actggcgatc ccgaccgcgt
      481 gcgcaattga aatgatccac acgatgtccc tgatgcacga tgatctgccg tgtatcgaca
      541 acgacgatct gcgtcgcggt aaaccgacta accacaaaat tttcggtgag gataccgcag
      601 tgactgctgg taacgcactg cactcttacg ccttcgagca tatcgcggtt tctacttcta
      661 aaaccgttgg tgctgaccgc atcctgcgta tggtgtccga gctgggtcgt gctactggct
      721 ctgaaggtgt tatgggtggt cagatggtag acatcgcatc cgaaggcgat ccgtctatcg
      781 acctgcagac cctggaatgg attcacatcc acaaaaccgc aatgctgctg gaatgctccg
      841 ttgtttgcgg tgcaatcatt ggcggtgcca gcgaaatcgt aatcgaacgt gcccgtcgct
      901 acgcccgctg tgttggtctg ctgttccagg tagttgatga cattctggac gtaactaaaa
      961 gcagcgacga actgggtaag actgcgggca aggacctgat ctctgataaa gccacctacc
     1021 caaagctgat gggtctggaa aaggccaagg agttctccga tgaactgctg aaccgtgcga
     1081 agggtgaact gtcctgcttc gacccagtta aagccgctcc gctgctgggc ctggcagact
     1141 acgtggcatt tcgtcagaat taacccgggg atcccgtttt aagaaggaga tatacatgcg
     1201 tcgcagtggt aattacaacc cgtcacgttg ggatgtgaat tttatccaat ccctgctgag
     1261 cgattacaaa gaggacaaac acgtcattcg ggcaagcgaa ctggttacac tggttaaaat
     1321 ggaactggaa aaagaaacag accaaattcg ccaattagag ctgattgacg acttacagcg
     1381 catggggctg agcgatcact ttcagaatga gttcaaagag attctgtcat ctatctatct
     1441 ggaccaccat tactataaaa acccttttcc gaaagaggaa cgtgatctgt atagcacaag
     1501 cctggccttt cggttactgc gtgagcatgg cttccaagta gcacaagagg tattcgatag
     1561 cttcaagaat gaagagggcg agttcaaaga atcactgtcc gatgatactc gtggcttatt
     1621 acaactgtat gaagcgagct ttctgctgac cgagggggag acgactctgg aaagcgcacg
     1681 ggaatttgcg accaaattct tagaagagaa agtaaatgaa ggcggcgtag atggcgatct
     1741 gttaacccgc atcgcgtatt ctctggacat tccgttacac tggcgtatta aacggccaaa
     1801 cgcgccggtg tggattgagt ggtatcgcaa acgtccagac atgaacccgg tggtgctgga
     1861 attagcaatt ttagacttaa acatcgtgca ggctcaattt caggaagaac tgaaagaatc
     1921 ttttcgctgg tggcgcaaca ccggcttcgt tgaaaaatta ccgtttgcgc gcgatcgtct
     1981 ggttgaatgt tatttctgga ataccggcat tatcgaaccg cgccagcacg cgagcgcccg
     2041 tattatgatg ggtaaggtca atgcgctgat tactgtcatt gacgatattt acgatgtata
     2101 cggcactctg gaagaactgg agcagtttac agatttaatc cgccggtggg acatcaatag
     2161 catcgaccag ctgccagatt acatgcagct gtgttttctg gcgctgaata actttgtgga
     2221 cgacacctct tatgacgtca tgaaagaaaa aggcgtgaat gttatccctt atctgcgtca
     2281 gagttgggtg gatctggccg ataaatatat ggttgaggca cgctggttct acggtgggca
     2341 caaaccgagt ttagaagaat atctggagaa ctcatggcag agtatcagtg ggccatgcat
     2401 gctgacgcat atcttctttc gcgtcacgga ttcatttacc aaggagacgg tcgattcctt
     2461 atataaatac catgacctgg ttcgttggtc atctttcgtc ctgcggctgg cggacgatct
     2521 gggtacgtct gtggaagagg tgtctcgtgg ggatgttccg aaatccttac agtgctacat
     2581 gtccgattat aacgcaagtg aagctgaagc tcgcaaacat gtgaagtggc tgatcgccga
     2641 agtgtggaag aagatgaacg ctgaacgtgt ttccaaggat agtccgtttg gtaaggattt
     2701 tattggttgc gccgttgatc tgggtcgtat ggcccagctg atgtatcata acggtgatgg
     2761 tcatgggacc cagcatccta ttattcatca gcagatgacg cgtaccctgt ttgaaccttt
     2821 cgccctcgag tgagtcgacc tgcaggcatg caagcttggc tgttttggcg gatgagagaa
     2881 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     2941 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     3001 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     3061 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3121 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     3181 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     3241 aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttttgtt tatttttcta
     3301 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3361 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3421 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3481 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3541 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3601 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3661 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3721 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3781 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3841 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3901 gcgtgacacc acgatgccta cagcaatggc aacaacgttg cgcaaactat taactggcga
     3961 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4021 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4081 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4141 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4201 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4261 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4321 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4381 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4441 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4501 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4561 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4621 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4681 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4741 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4801 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4861 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4921 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4981 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5041 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5101 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5161 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5221 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5281 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     5341 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5401 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     5461 cagcagatca attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac accatcgaat
     5521 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     5581 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     5641 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     5701 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     5761 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     5821 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     5881 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     5941 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     6001 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     6061 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     6121 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     6181 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     6241 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     6301 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     6361 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     6421 gctcatgtta tatcccgccg tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     6481 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     6541 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     6601 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6661 ggcagtgagc gcaacgcaat taatgtgagt tagcgcgaat tgat
//