LOCUS       pTpal-fdc                7837 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses A. thaliana PAL2 and S. cerevisiae FDC1 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pTpal-fdc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7837)
  AUTHORS   McKenna R, Pugh S, Thompson B, Nielsen DR
  TITLE     Microbial production of the aromatic building-blocks (S)-styrene 
            oxide and (R)-1,2-phenylethanediol from renewable resources.
  JOURNAL   Biotechnol J. 2013 Dec;8(12):1465-75. doi: 10.1002/biot.201300035. 
            Epub 2013 Jul 29.
  PUBMED    23801570
REFERENCE   2  (bases 1 to 7837)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7837
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4020..4037)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4190..4276
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4368..4395
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4415..4506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4507..5367
                     /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(4725..4744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5538..6126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6027..6046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6280..6297
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6312..6452
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6378..6397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6538..6560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6638..6715
                     /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             6716..7798
                     /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(6735..6754)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg gcgggaggta accaatggat caaatcgaag
      301 caatgttgtg cggcggagga gagaagacaa aagtggcggt tactacgaag actttggcag
      361 atccattgaa ttggggttta gcagcggatc aaatgaaagg aagtcattta gatgaagtga
      421 agaagatggt cgaagagtat cgtagaccag tcgtgaatct tggcggagaa acactgacga
      481 tcggacaagt tgctgccatc tccaccgtag gaggcagcgt taaggttgag ttagcggaga
      541 cttcaagagc cggtgtgaaa gctagcagtg attgggttat ggagagcatg aacaaaggta
      601 ctgacagtta cggagtcacc accggctttg gtgctacttc tcaccggaga accaaaaacg
      661 gcaccgcatt acaaacagaa ctcattagat ttttgaacgc cggaatattc ggaaacacga
      721 aggagacatg tcacacactg ccgcaatccg ccacaagagc cgccatgctc gtcagagtca
      781 acactcttct ccaaggatac tccgggatcc gattcgagat cctcgaagcg attacaagtc
      841 tcctcaacca caacatctct ccgtcactac ctctccgtgg aaccattacc gcctccggcg
      901 atctcgttcc tctctcttac atcgccggac ttctcaccgg ccgtcctaat tccaaagcca
      961 ccggtcccga cggtgaatcg ctaaccgcga aagaagcttt tgagaaagcc ggaatcagta
     1021 ctggattctt cgatttacaa cctaaggaag gtttagctct cgttaatggc acggcggttg
     1081 gatctggaat ggcgtcgatg gttctattcg aagcgaatgt ccaagcggtg ttagcggagg
     1141 ttttatcagc gatcttcgcg gaggttatga gcgggaaacc tgagtttacc gatcatctga
     1201 ctcatcgttt aaaacatcat cccggacaaa tcgaagcggc ggcgataatg gagcacatac
     1261 tcgacggaag ctcatacatg aaattagctc aaaaggttca cgagatggat ccattgcaga
     1321 aaccaaaaca agatcgttac gctcttcgta catctcctca atggctaggt cctcaaattg
     1381 aagtaatccg tcaagctacg aaatcgatag agcgtgaaat caactccgtt aacgataatc
     1441 cgttgatcga tgtttcgagg aacaaggcga ttcacggtgg taacttccaa ggaacaccaa
     1501 tcggagtttc tatggataac acgagattgg cgattgctgc gattgggaag ctaatgtttg
     1561 ctcaattctc tgagcttgtt aatgatttct acaacaatgg acttccttcg aatctaactg
     1621 cttcgagtaa tccaagtttg gattatggat tcaaaggagc agagattgct atggcttctt
     1681 attgttctga gcttcaatac ttggctaatc cagtcacaag ccatgttcaa tcagctgagc
     1741 aacataatca agatgtgaac tctcttggtt tgatctcgtc tcgtaaaaca tctgaagctg
     1801 tggatattct taagctaatg tcaacaacgt tccttgtggg gatatgtcaa gctgttgatt
     1861 tgagacattt ggaggagaat ctgagacaaa ctgtgaagaa cacagtttct caagttgcta
     1921 agaaagtgtt aaccactgga atcaacggtg agttacatcc gtcaaggttt tgcgagaagg
     1981 acttgcttaa ggttgttgat cgtgagcaag tgttcacgta tgtggatgat ccttgtagcg
     2041 ctacgtaccc gttgatgcag agactaagac aagttattgt tgatcacgct ttgtccaacg
     2101 gtgagactga gaagaatgca gtgacttcga tctttcaaaa gattggagct tttgaagagg
     2161 agcttaaggc tgtgcttcca aaggaagttg aagcggctag agcggcttat gggaatggaa
     2221 ctgcgccgat tcctaaccgg attaaggaat gtaggtcgta tccgttgtat aggttcgtga
     2281 gggaagagct tggaacgaag ttgttgactg gagaaaaggt tgtgtctccg ggagaggagt
     2341 ttgataaggt cttcactgct atgtgtgaag gtaaacttat tgatccgttg atggattgtc
     2401 tcaaggaatg gaacggagct ccgattccga tttgctaatc tagagtcgac ctgcaggggg
     2461 aggaattata tgaggaagct aaatccagct ttagaattta gagactttat ccaggtctta
     2521 aaagatgaag atgacttaat cgaaattacc gaagagattg atccaaatct cgaagtaggt
     2581 gcaattatga ggaaggccta tgaatcccac ttaccagccc cgttatttaa aaatctcaaa
     2641 ggtgcttcga aggatctttt cagcatttta ggttgcccag ccggtttgag aagtaaggag
     2701 aaaggagatc atggtagaat tgcccatcat ctggggctcg acccaaaaac aactatcaag
     2761 gaaatcatag attatttgct ggagtgtaag gagaaggaac ctctcccccc aatcactgtt
     2821 cctgtgtcat ctgcaccttg taaaacacat atactttctg aagaaaaaat acatctacaa
     2881 agcctgccaa caccatatct acatgtttca gacggtggca agtacttaca aacgtacgga
     2941 atgtggattc ttcaaactcc agataaaaaa tggactaatt ggtcaattgc tagaggtatg
     3001 gttgtagatg acaagcatat cactggtctg gtaattaaac cacaacatat tagacaaatt
     3061 gctgactctt gggcagcaat tggaaaagca aatgaaattc ctttcgcgtt atgttttggc
     3121 gttcccccag cagctatttt agttagttcc atgccaattc ctgaaggtgt ttctgaatcg
     3181 gattatgttg gcgcaatctt gggtgagtcg gttccagtag taaaatgtga gaccaacgat
     3241 ttaatggttc ctgcaacgag tgagatggta tttgagggta ctttgtcctt aacagataca
     3301 catctggaag gcccatttgg tgagatgcat ggatatgttt tcaaaagcca aggtcatcct
     3361 tgtccattgt acactgtcaa ggctatgagt tacagagaca atgctattct acctgtttcg
     3421 aaccccggtc tttgtacgga tgagacacat accttgattg gttcactagt ggctactgag
     3481 gccaaggagc tggctattga atctggcttg ccaattctgg atgcctttat gccttatgag
     3541 gctcaggctc tttggcttat cttaaaggtg gatttgaaag ggctgcaagc attgaagaca
     3601 acgcctgaag aattttgtaa gaaggtaggt gatatttact ttaggacaaa agttggtttt
     3661 atagtccatg aaataatttt ggtggcagat gatatcgaca tatttaactt caaagaagtc
     3721 atctgggcct acgttacaag acatacacct gttgcagatc agatggcttt tgatgatgtc
     3781 acttcttttc ctttggctcc ctttgtttcg cagtcatcca gaagtaagac tatgaaaggt
     3841 ggaaagtgcg ttactaattg catatttaga cagcaatatg agcgcagttt tgactacata
     3901 acttgtaatt ttgaaaaggg atatccaaaa ggattagttg acaaagtaaa tgaaaattgg
     3961 aaaaggtacg gatataaata aaagcttggc tgttttggcg gatgagagaa gattttcagc
     4021 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     4081 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     4141 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     4201 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4261 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     4321 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     4381 gacggatggc ctttttgcgt ttctacaaac tctttttgtt tatttttcta aatacattca
     4441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4741 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5041 acgatgccta cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6301 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6361 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     6421 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     6481 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6541 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca
     6601 attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac
     6661 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     6721 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     6781 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     6841 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     6901 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     6961 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     7021 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     7081 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     7141 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     7201 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     7261 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     7321 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     7381 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     7441 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     7501 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     7561 tatcccgccg tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     7621 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     7681 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     7741 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7801 gcaacgcaat taatgtgagt tagcgcgaat tgatctg
//