LOCUS       sequence-371218-        5713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of S. coelicolor butyrolactone biosynthesis genes in E. 
            coli. Heterologous production of SCBs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5713)
  AUTHORS   Biarnes-Carrera M, Lee CK, Nihira T, Breitling R, Takano E
  TITLE     Orthogonal Regulatory Circuits for Escherichia coli Based on the 
            gamma-Butyrolactone System of Streptomyces coelicolor.
  JOURNAL   ACS Synth Biol. 2018 Apr 20;7(4):1043-1055. doi: 
            10.1021/acssynbio.7b00425. Epub 2018 Mar 19.
  PUBMED    29510026
REFERENCE   2  (bases 1 to 5713)
  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"; doi: 
            "10.1021/acssynbio.7b00425"; journalName: "ACS Synth Biol"; date: 
            "2018-04-20- 20"; volume: "7"; issue: "4"; pages: "1043-1055"
COMMENT     Sequence Label: pTE1059
FEATURES             Location/Qualifiers
     source          1..5713
                     /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"
     CDS             754..771
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             772..789
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     protein_bind    1781..1799
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1840..1862
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2655..2672
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2721..2739
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3671..3688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3723..3794
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3810..3837
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3865..3882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3999..4544)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4079..4098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4658..4752)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4783..5577)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4894..4913)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5504..5523
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
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 tcaaaagatc
      721 ttttaagaag gagatataca tatgagagga tcgcatcacc atcaccatca cctggtgccg
      781 cgtggcagcc ccgaagcagt agttttgatc aattctgcgt ccgatgccaa ctcgatcgaa
      841 caaaccgcat tgccggttcc gatggcgctt gtccaccgga ccagggtgca ggacgcgttc
      901 ccggtcagct ggataccgaa gggcggtgac cggttctcgg tcaccgccgt cctgccccac
      961 gaccacccgt tcttcgcacc ggtccacggg gaccgacacg atccgctgct gatcgccgag
     1021 accctgcgtc aggcggcgat gctcgtcttc cacgccggct acggcgtgcc ggtgggctac
     1081 cacttcctga tggccacgct ggactacacc tgccacctcg accacctcgg cgtgtcgggc
     1141 gaggtcgcgg agctggaggt ggaagtggcc tgttcccagc tgaagttccg cggcgggcag
     1201 cccgtacagg gacaggtgga ctgggccgtg cgccgcgccg gacggctcgc tgccacgggg
     1261 actgccacga cgcgcttcac cagtcctcaa gtctaccggc ggatgcgcgg cgacttcgcg
     1321 actcccaccg catcggtgcc cgggaccgcg cccgtgcccg cggcgcgcgc cggtcgcacc
     1381 cgcgacgagg acgtggtcct gtcggcgagt tcgcagcagg acacgtggcg actgcgggtg
     1441 gacaccagtc acccgaccct cttccagcgc cccaacgacc acgtaccggg catgctgctg
     1501 ctcgaggcgg cacggcaggc ggcgtgcctc gtgaccggtc cggcgccctt cgtgccgtcg
     1561 atcggcggca cccggttcgt ccggtacgcg gagttcgaca gcccgtgctg gatccaggcg
     1621 acggtccggc cggggccggc ggcggggctg accaccgtgc gggtcaccgg gcatcaggac
     1681 ggcagcctcg tcttcctcac cacgctgtcc ggccccgcgt tctccggctg aagcttacaa
     1741 cgtcgtgtta gctgcctttc gtcttcaata attcttgaca tccctatcag tgatagagat
     1801 acttcgtgca atttttatat caccgccagg ggtacaactt ttgtttaact ttaagaagga
     1861 gatataccat gcgtgcacat ggcacccgtt atggtcgtcc gctggagggc aagaccgccc
     1921 tggtgaccgg gggaagccgg ggcatcggac gcggtatcgc cctgcggctg gccgcggacg
     1981 gggcgctcgt ggcagtccac tacggcagca gcgaggcggc ggcgcgagag accgtcgaga
     2041 ccatccggag cagcggcgga caggcgttgg ccatccgggc cgaactcggc gtcgtgggtg
     2101 acgcggccgc gctctacgcg gcgttcgacg ccgggatggg cgagttcgga gtgccgcccg
     2161 agttcgacat cctggtgaac aacgcgggcg tcagcggctc gggacggatc acggaagtca
     2221 ccgaggaggt cttcgaccga ctggtcgccg tcaacgtccg ggcaccgctg ttcctggtcc
     2281 agcacggact gaaacgactg cgcgacggcg ggcggatcat caacatctcg tctgccgcga
     2341 cccggcgcgc gttccccgag tccatcggat acgcgatgac caagggtgcg gtggacacgc
     2401 tcacgctcgc cctggccagg caactggggg aacgggggat cacggtcaac gcggtggcgc
     2461 cgggtttcgt ggagacggac atgaacgcac ggcggcggca gacccccgag gcggccgcgg
     2521 cactggccgc ctactccgtg ttcaaccgca tcggcagacc cgacgacatc gccgacgtgg
     2581 tggcctttct ggcctccgac gactcacggt ggatcaccgg ccagtacgtc gatgccacag
     2641 gcggaacgat tctccatcat caccatcacc actgagtaac aaaagtgtct ataatcacgg
     2701 cagaaaagtc cacattgaca tccctatcag tgatagagat acattccacc cctatcagtg
     2761 atagagagcg ttacccaaca acttcatata cctataaaac aaaggagaaa taatgggtac
     2821 cattcttgtg accggcgtga ccggtgtggt cggccgtcag gtcgcccacc tgctggcccg
     2881 gaccggtccc gtcagagtcc tcgcccggcg gccggaacgg gtgaccgtca ccggcccgaa
     2941 cgtcgaggtc tgcgcgggcg agtacgccga ccgcccgggc ctggaccggg ccctcaacgg
     3001 ggtgcgggcg gcgttcctcg tcacgaacag tgccaccgag ccggacgacg agcgtttcgc
     3061 ggcggcggcg cgcgaggccg gcgtccggca cctggtgaag ctgtccatgc tcgcggtggg
     3121 ggaaccgggc gcggacgact tcatcacgcg gcggcagcgc gagaacgagc gggccgtacg
     3181 cgagtccggg ctcgactgga ccttcctgcg ggccaggacc ttcatgtcca acacgctgtc
     3241 ctgggcgccc gccatccggt ccgacggcgt ggtccgcgcg ctgtacggca cgtccccggt
     3301 tgcctgcgtc gatccgcgcg acatcgcgga agtggcggtc agggcactga cccggcccgg
     3361 ccacgaggga cgggcgtacg cgctgtccgg gcccgaggcg atcaccgcca ggcagcagac
     3421 ggcgcggctc tccgaggtgc tggggcggtc actgcgtttc gaggagctgg ggctggaggc
     3481 ggcgcgcgag cagctgctcc ggaagtatcc gaggcccgtc gccgacgcgt tcctggagag
     3541 cgccgagcgg cagcgggcgg gggccaaggc gcaggtcgtg ccgaccgtgc gggaagtgac
     3601 gggccggccc gccaggccgt tccgcatctg ggcggcggac cacgcggaag ccttcggccg
     3661 aagccagggg catcatcacc atcaccactg agatccaaac tcgagtaagg atctccaggc
     3721 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3781 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     3841 atacctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     3901 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     3961 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
     4021 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     4081 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     4141 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     4201 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     4321 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     4381 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     4441 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     4501 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     4561 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     4621 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     4681 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     4741 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
     4801 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     4861 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     4921 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     4981 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     5041 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     5101 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     5161 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     5221 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     5281 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     5341 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     5401 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     5461 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     5521 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     5581 aaacgatcct catcctgtct cttgatcaga tcatgatccc ctgcgccatc agatccttgg
     5641 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     5701 agctggcaat tcc
//
