LOCUS       sequence-413670-        7487 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Production of labda-7,13(16),14-triene in E. coli MEV20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Lin GM, Voigt CA
  TITLE     Design of a redox-proficient Escherichia coli for screening 
            terpenoids and modifying cytochrome P450s
  JOURNAL   Nature Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5
REFERENCE   2  (bases 1 to 7487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature 
            Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5"
COMMENT     Sequence Label: pLRD10
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(90..133)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    136..157
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        238..256
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    257..281
                     /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)."
     promoter        2708..2726
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2727..2751
                     /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)."
     misc_feature    4035..4055
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4056..4113
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(4137..4154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4688..4910
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4958..5908
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFWIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(6522..7313,7314..7382))
                     /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     7145..7164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7383..7487)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
      121 taaggatgat ttctggaatt cgcggccgct tctagagcgc gaagacttaa tcaaatccgt
      181 agcagtgcac aagcaagtga gtgctgacga gctcaggagc cgcaatacac agtgtgctaa
      241 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctaaac gccagccact
      301 aaagcggtca acgcatgtgc tttgcgttct gatgagacag tgatgtcgaa accgcctcta
      361 caaataattt tgtttaagat aaattactct aaggagggct gatgtgcaaa gctatttata
      421 accccgaaaa aggtctgtta aaacactcgg acctgcagct ggaagatgat aaatatagcg
      481 acctgcagtt ggtggatgaa aaacacacta ttaaacgctc agtagaatct atcaagtccg
      541 cactgtccct ggtggatgac ggcgatctga cgctgtctgc gtatgacacg gcctgggtcg
      601 ccctgatcga ggacgtcaat ggtagaagcg gtttcccgca attccccagc tgcctgcaat
      661 ggatcgtttc acagcagctg cccgacggct catggggtga accactgatg ttcctggctt
      721 ttgaccgcct gctgaacacg ctggctagtg tggttgcact gacaaaatgg aacattcgcc
      781 cggatatttg tcagaaaggc atgaaatatg tgctggaaaa cctgaataag ctcgttgacg
      841 aaaaagaaga acacatgacc cccggcttcg agttgctgtt cccgaaactc atcgaactgg
      901 cacagaaact ggacatcaag atgcccatgg acagcccagc gttaaaagaa ttatatgcgc
      961 gtcgtgatac gaaactggcc aaaatcccga agaaaatttt ccataagatg ccgacgatcc
     1021 tcctctacag cttagagggg atgaacgatc tcgaatggga taaattgctg aaactgaaat
     1081 cagagaacgg ctccttcctg tgttcacccg cggccacagc ttttgcattt atggagacaa
     1141 aagaccaaga ttgcctcgcg tacctgaccg atcttgttgc gaagtttaat ggtggcgttc
     1201 cgacatttta tccaaccgat atgtacgaac agatttggat tgttgatcgt ttgcaaagac
     1261 tggggattgc acattatttc tcttcagaga tcaacaactt cgtcgaccac atctatcgct
     1321 attgggatca gaagggaatc agttttgcca gaaaatgtaa tcttccggac attgacgaca
     1381 ccgctatggg tttccgtgta ctgcgtaccc atgggtacca ggtatcgagc gatgtgttcc
     1441 agcatttcga aaaagatggc caattttact gctattgggg ccagacagcg gaagccgtga
     1501 ccgtcatgtt taacctgtac cgtgccagcc aagttttgtt cccaggcgaa aagattctgg
     1561 ataatgctaa aaagtttgcg cacaactttc ttaccgagaa ggttgcgacc aatcaggttt
     1621 tcgataagtg gattattacc aaagacattc tgggcgaagt gcaatatgcg ttagatgtcc
     1681 cgtggtatgc aagcctgccg cgtctggaag ctagatatta cttagatcag tatgccgggg
     1741 acggcgatgt ttggatcgcg aaaacccttt accgcctcaa atatgtatcg aacaacgaat
     1801 atctcgaaac ggctaagctg gattacaatc attgtcagaa gattcacaag ctggagtgga
     1861 gctatataca aaaatggttc ctggatctca aaatcgaaga aagcatcaac actcgtacgc
     1921 tttggagtta ttaccaggcc gcggcgagta tcttccaccc tgaaagatac aacgaacgtc
     1981 tggcctgggc gaaaacgaat gtcctggttg acacgataac caccttcttt agcaagcaac
     2041 aaatgagtaa agatgatatt cagggctttg tgaatcagct gactaacacc gaaaaccagc
     2101 agacctacgg caaaatgagt cacatgttaa tcgatgcgct gaatgaaacg ctgaagcaca
     2161 tcagtatgaa ggcccgcgaa acccatggca ttgatatcta cccacatctg cagtccagct
     2221 ggaagaaatg gttactgtct tgtatgaacg ggcctaatgt tgccggcgtg gcggagctga
     2281 ttgttgaaac cataaatctt acttcaggcc gcagcttctc caatgattta ctttcgcatc
     2341 cccaatacaa acaaattacg tctataacca acgacctttg tcatcagctg tgttcaaaag
     2401 gcaaccgtgc gatcgggtcc gaaattgaat ctaaaatgca ggaactggtc caactggtgt
     2461 tttcggactc ctccgacggc ctcgatcccg atgtgaagaa aacctacctc gttgtggcca
     2521 aatcctttta ttatatggca tactttgacg ccaaaacaat cgattcccac atcaacaaag
     2581 tgctgtttga aatggtggtt taataattat gccttccgct aatcaactcg gtaccaaatt
     2641 ccagaaaaga gacgctttcg agcgtctttt ttcgttttgg tccgtggaca agcacagcat
     2701 agccaagtaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctaggt
     2761 ccgaagatac ttagggtgtc tcaaggtgcg taccttgact gatgagtccg aaaggacgaa
     2821 acacccctct acaaataatt ttgtttaaga ggcttagcta ataggaggta gatatgggac
     2881 gtagagcacg aagcgcccgc agttttggtg tttcgccttt gtggggcggc gtcagcgtgc
     2941 gctctgggga tagaggtgaa gcggcagtcg gcggtttgtg ggaggtgcct gatttttggg
     3001 gcctgtttcc gtcacggatt tcgccactcg ccggcgaagt ggaaagcggg acccgcgtct
     3061 ggcttgatgg atggcgtctg gtagaagaag ctggccctgg agaacgtctg aaggcgtcga
     3121 aagtggggcg tcttgtcgcg cttgcgtatc cggatgcgcc cgcggatctg cttcgttggg
     3181 cggcagacct gtttgcctgg cttactgcgt ttgatgatgt ccatgttgaa gcaccgggag
     3241 ttacgacagc cgaactggga ccgcacatgg cgtctttcgt tggcgttttg gaaacaggca
     3301 ccgccccggg agcggcaccc acaccatttc cagctgcgct ggcggaactg ttggatcgcg
     3361 cgcgcgaatt actgacccct ctgcaagaag aacgcgtgcg tgctcggttg ggtaaagtgt
     3421 ttgtggcaat gctctgggaa attacgactc gtgagcgtac cgtatcaacc gcagaatacg
     3481 aaacgatgcg ccctcataca tttttctcag cggttggcgc tgcccttgtc gagccctgtg
     3541 ccggtctgga cctgagtcat ggtgtccgcg ccgacccagg ggttcgccgc ctgacccaag
     3601 cattagctac attgtgggaa cgtacgaatg atctgtatag cttcgcgtac gaacaacgtg
     3661 cattaggctc tgtcccccgg acattgccgt ggctcatcgc ccaagaacgc ggtctgcccc
     3721 tggatgctgc atttgctgaa gctggacgct ggtgcgaaga agaagctgtc ctggctcatc
     3781 ggctgatcgg tgaactgagc gccagcgcac gcgaaggtgt acctgaatac gccggtgcgg
     3841 ttgcgcacgc cattggtggt acgcgccgcc tgtatgaagt gagtgatcgc tggcgtgaag
     3901 agtaataagc ctcgttgctc ttcataaatc atccctatga ccccttcaaa aaaaaaaaac
     3961 accctaacgg gtgttttttt ttttttggtc tgcctctcgc tgacggtcac aatgcgctta
     4021 cgaaagtctt cctgtactag tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca
     4081 ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgttatgca ggcttcctcg
     4141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4201 gcggtaatga cagtaagacg ggtaagcctg ttgatgatac cgctgcctta ctgggtgcat
     4261 tagccagtct gaatgacctg tcacgggata atccgaagtg gtcagactgg aaaatcagag
     4321 ggcaggaact gctgaacagc aaaaagtcag atagcaccac atagcagacc cgccataaaa
     4381 cgccctgaga agcccgtgac gggcttttct tgtattatgg gtagtttcct tgcatgaatc
     4441 cataaaaggc gcctgtagtg ccatttaccc ccattcactg ccagagccgt gagcgcagcg
     4501 aactgaatgt cacgaaaaag acagcgactc aggtgcctga tggtcggaga caaaaggaat
     4561 attcagcgat ttgcccgagc ttgcgagggt gctacttaag cctttagggt tttaaggtct
     4621 gttttgtaga ggagcaaaca gcgtttgcga catccttttg taatactgcg gaactgacta
     4681 aagtagtgag ttatacacag ggctgggatc tattcttttt atcttttttt attctttctt
     4741 tattctataa attataacca cttgaatata aacaaaaaaa acacacaaag gtctagcgga
     4801 atttacagag ggtctagcag aatttacaag ttttccagca aaggtctagc agaatttaca
     4861 gatacccaca actcaaagga aaaggactag taattatcat tgactagccc atctcaattg
     4921 gtatagtgat taaaatcacc tagaccaatt gagatgtatg tctgaattag ttgttttcaa
     4981 agcaaatgaa ctagcgatta gtcgctatga cttaacggag catgaaacca agctaatttt
     5041 atgctgtgtg gcactactca accccacgat tgaaaaccct acaaggaaag aacggacggt
     5101 atcgttcact tataaccaat acgctcagat gatgaacatc agtagggaaa atgcttatgg
     5161 tgtattagct aaagcaacca gagagctgat gacgagaact gtggaaatca ggaatccttt
     5221 ggttaaaggc ttttggattt tccagtggac aaactatgcc aagttctcaa gcgaaaaatt
     5281 agaattagtt tttagtgaag agatattgcc ttatcttttc cagttaaaaa aattcataaa
     5341 atataatctg gaacatgtta agtcttttga aaacaaatac tctatgagga tttatgagtg
     5401 gttattaaaa gaactaacac aaaagaaaac tcacaaggca aatatagaga ttagccttga
     5461 tgaatttaag ttcatgttaa tgcttgaaaa taactaccat gagtttaaaa ggcttaacca
     5521 atgggttttg aaaccaataa gtaaagattt aaacacttac agcaatatga aattggtggt
     5581 tgataagcga ggccgcccga ctgatacgtt gattttccaa gttgaactag atagacaaat
     5641 ggatctcgta accgaacttg agaacaacca gataaaaatg aatggtgaca aaataccaac
     5701 aaccattaca tcagattcct acctacataa cggactaaga aaaacactac acgatgcttt
     5761 aactgcaaaa attcagctca ccagttttga ggcaaaattt ttgagtgaca tgcaaagtaa
     5821 gtatgatctc aatggttcgt tctcatggct cacgcaaaaa caacgaacca cactagagaa
     5881 catactggct aaatacggaa ggatctgagg ttcttatggc tcttgtatct atcagtgaag
     5941 catcaagact aacaaacaaa agtagaacaa ctgttcaccg ttacatatca aagggaaaac
     6001 tgtccatatg cacagatgaa aacggtgtaa aaaagataga tacatcagag cttttacgag
     6061 tttttggtgc attcaaagct gttcaccatg aacagatcga caatgtaaca gatgaacagc
     6121 atgtaacacc taatagaaca ggtgaaacca gtaaaacaaa gcaactagaa catgaaattg
     6181 aacacctgag acaacttgtt acagctcaac agtcacacat agacagcctg aaacaggcga
     6241 tgctgcttat cgaatcaaag ctgccgacaa cacgggagcc agtgacgcct cccgtgggga
     6301 aaaaatcatg gcaattctgg aagaaatagc gctttcagcc ggcaaaccgg ctgaagccgg
     6361 atctgcgatt ctgataacaa actagcaaca ccagaacagc ccgtttgcgg gcagcaaaac
     6421 ccgtaccgat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6661 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat ataacccact
     7261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcg
//
