LOCUS       sequence-456749-        7916 bp    DNA     circular SYN 15-APR-2025
DEFINITION  An IPTG-inducible promoter controlling expression of eutR; PEutS, 
            extended controlling expression of sfgfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7916)
  AUTHORS   McNerney MP, Piorino F, Michel CL, Styczynski MP
  TITLE     Active Analyte Import Improves the Dynamic Range and Sensitivity of 
            a Vitamin B(12) Biosensor.
  JOURNAL   ACS Synth Biol. 2020 Feb 21;9(2):402-411. doi: 
            10.1021/acssynbio.9b00429. Epub 2020 Feb 13.
  PUBMED    31977200
REFERENCE   2  (bases 1 to 7916)
  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"; doi: 
            "10.1021/acssynbio.9b00429"; journalName: "ACS Synth Biol"; date: 
            "2020-02-21- 21"; volume: "9"; issue: "2"; pages: "402-411"
COMMENT     Sequence Label: pEutRwc
FEATURES             Location/Qualifiers
     source          1..7916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(403..408,409..425,426..431)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             479..1669
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(831..850)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      complement(1799..1828)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1875..1896
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             1971..3053
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1990..2009)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3073..3144
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3160..3187
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    3204..3220
                     /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)."
     protein_bind    3227..3243
                     /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     complement(5874..5902)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6497..6517
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    6561..6581
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      6617..6648
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      7159..7703
                     /direction=RIGHT
                     /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     7604..7623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7820..7837
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gcgctgaggt
       61 ctgcctcgtg aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc
      121 cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt
      181 ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc
      241 ttcaactcag caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga
      301 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
      361 aacagtaata caaggggtgt ttactagagg agattctcat gtttgacagc ttatcatcga
      421 taagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat
      481 gaaatctaac aatgcgctca tcgtcattct cggcaccgtc accctggacg ctgtaggcat
      541 aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagtat
      601 tgccagtcac tatggcgtgc tgcttgcgct ctatgcgttg atgcaatttc tttgcgcacc
      661 cgttctcgga gccctgtccg accgctttgg ccgccgtcca gtcctgctcg cttcgctcct
      721 tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga ttctctacgc
      781 cggacgcatc gtggcgggca tcacgggtgc cacaggtgcg gttgctggtg cctatatcgc
      841 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      901 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggtgccatct ccttgcatgc
      961 accattcctt gcggcggcgg tgctcaacgg cctcaacctc ctcctgggct gcttccttat
     1021 gcaggaatcg cataagggag agcgccgtcc gatgcccttg cgtgccttca atccagtcag
     1081 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg ttttctttat
     1141 catgcaactc gtaggacagg ttccggcagc gctctgggtc attttcggcg aggaccgctt
     1201 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     1261 cgctcaagcc ttcgtcacgg gccccgccac caaacgtttc ggcgagaagc aggccattat
     1321 cgcgggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgcggctg
     1381 gatggccttc cccattatga ttcttctcgc ttccggcggc atcggtatgc ccgcgttgca
     1441 ggccatgctg tcccgccaag tagatgacga ccatcaggga cagcttcaag ggtcgctcgc
     1501 ggctcttacc agcctcactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     1561 ctcggcgagc acatggaacg ggttggcatg gattgtaggt gccgcccttt accttgtctg
     1621 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctaat aatactagct
     1681 ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
     1741 tacttcgcgt ttgccacctg acgtctaaga aaaggaatat tcagcaattt gcccgtgccg
     1801 aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gctacgtaat tagttagtta
     1861 gcccttagtg actcgaattc gcggccgctt ctagagtaac accgtgcgtg ttgactattt
     1921 tacctctggc ggtgataatg gttgctacta gagtcacaca ggactactag atgaaaccag
     1981 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     2041 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     2101 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     2161 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     2221 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     2281 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     2341 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     2401 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2461 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2521 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2581 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2641 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     2701 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     2761 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     2821 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     2881 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     2941 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3001 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgataatact
     3061 agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3121 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     3181 ttctgcgttt atatactaga gaattgtgag cggataacaa ttgacattgt gagcggataa
     3241 caagatactg agcacacttc gccgaagcca ttttaattcc acaacttatg aaaaagaccc
     3301 gtacagccaa tttgcaccat ctttatcatg aacccttacc cgaaaacctg aagctcacgc
     3361 cgaaggtcga agtggataat gttcatcaac gacagacaac ggatgtctat gaacatgctt
     3421 taacgattac cgcctggcag cagatttacg atcagctgca tccgggcaag tttcatggtg
     3481 aatttacgga aattctactc gatgatattc aggtttttcg tgaatacacc ggtctggcgc
     3541 tgcgtcagtc gtgcctggtc tggccgaact cgttctggtt tggcattccg gcgacgcgcg
     3601 gtgagcaggg atttatcggt tcgcaatgtc tgggaagcgc ggaaatcgcc acccgccctg
     3661 gtggcactga atttgaactg agcacgccgg atgattacac gatcctgggc gtggtgcttt
     3721 ctgaagatgt catcacccgg caggctaact ttttgcataa cccggatcgg gtattacata
     3781 tgttgcgtaa ccagtcggcg ctggaagtga aagagcagca taaagccgcg ctgtggggct
     3841 ttgtccaaca ggcgctggcg acgttttgcg agaatccgga aaatctccat cagcctgcag
     3901 tgcgaaaagt gctgggggat aatttgctaa tggcgatggg ggccatgctg gaagaagcgc
     3961 aaccaatggt gacggcggaa agcatcagtc atcagagtta ccgtcgattg ctttcccgcg
     4021 cccgtgaata tgtgctggaa aacatgtccg aaccggtgac ggtgctggat ttgtgtaatc
     4081 aactgcatgt cagccgccgc acgctacaaa acgcgtttca cgctatttta ggcattggcc
     4141 cgaacgcgtg gctgaaacgc attcgcctga acgccgtacg ccgcgaactg ataagtccgt
     4201 ggtcgcaaag tatgacggta aaagacgccg ccatgcagtg gggattctgg catctggggc
     4261 aatttgccac ggattaccag cagctgtttt ccgagaagcc gtcactgacg ctgcatcagc
     4321 ggatgcggga gtgggggtga aacagagcga agtggtggct tcagcgtatg gcgatcagga
     4381 tctgagcttt ggtccggaat acatcattcc aaaaccgttt gatccgcgct tgatcgttaa
     4441 gatcgctcct gcggtcgcta aagccgcgat ggagtcgggc gtggcgactc gtccgattgc
     4501 tgatttcgac gtctacatcg acaagctgac tgagttcgtt tacaaaacca acctgtttat
     4561 gaagccgatt ttctcccagg ctcgcaaagc gccgaagcgc gttgttctgc cggaagggga
     4621 agaggcgcgc gttctgcatg ccactcagga actggtaacg ctgggactgg cgaaaccgat
     4681 ccttatcggt cgtccgaacg tgatcgaaat gcgcattcag aaactgggct tgcagatcaa
     4741 agcgagcgtt gattttgaga tcgtcaataa cgaatccgat ccgcgcttta aagagtactg
     4801 gaccgaatac ttccagatca tgaagcgtcg cggcgtcact caggaacagg cgcagcgggc
     4861 gctgatcagt aacccgacag tgatcggcgc gatcatggtt cagcgtgggg aagccgatgc
     4921 aatgatttgc ggtacggtgg gtgattatca tgaacatttt agcgtggtga aaaatgtctt
     4981 tggttatcgc gatggcgttc acaccgcagg tgccatgaac gcgctgctgc tgccgagtgg
     5041 taacaccttt attgccgata catatgttaa tgatgaaccg gatgcagaag agctggcgga
     5101 gatcaccttg atggcggcag aaactgtccg tcgttttggt attgagccgc gcgttgcttt
     5161 gttgtcgcac tccaactttg gttcttctga ctgcccgtcg tcgagcaaaa tgcgtcaggc
     5221 gctggaactg gtcagggaac gtgcaccaga actgatgatt gatggtgaaa tgcacggcga
     5281 tgcagcgctg gtggaagcga ttcgcaacga ccgtatgccg gacagctctt tgaaaggttc
     5341 cgccaatatt ctggtgatgc cgaacatgga agctgcccgc attagttaca acttactgcg
     5401 tgtttccagc tcggaaggtg tgactgtcgg cccggtgctg atgggtgtgg cgaaaccggt
     5461 tcacgtgtta acgccgatcg catcggtgcg tcgtatcgtc aacatggtgg cgctggccgt
     5521 ggtagaagcg caaacccaac cgctgtaatt ttttttaact ctcacgctta tcctgaatat
     5581 tcagggtaag cagtttagct gcaatatatt agtaaagctt attactgagt ttgcgaataa
     5641 taaaaaaaag cagtctatat aatatctcga tattatttta tttatattca tgcgttgcat
     5701 atgaaagttt atgcaccaca gcgaatatct ctccattctt agtgatctac ctcacctttt
     5761 aaacgcgctt gccgaatttt gttatttact ctgacgaaaa attgtcacga tacacgaaag
     5821 tttttcacag gcggcgactc atggctagca aaggagaaga actcttcact ggagttgtcc
     5881 caattcttgt tgaattagat ggtgatgtta acggccacaa gttctctgtc agtggagagg
     5941 gtgaaggtga tgcaacatac ggaaaactta ccctgaagtt catctgcact actggcaaac
     6001 tgcctgttcc atggccaacc ctggtcacta ctctgtgcta tggtgttcaa tgcttttcaa
     6061 gatacccgga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     6121 tacaggaaag gaccatcttc ttcaaagatg acggcaacta caagacacgt gctgaagtca
     6181 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgac ttcaaggaag
     6241 atggcaacat tctgggacac aaattggaat acaactataa ctcacacaat gtatacatca
     6301 tggcagacaa acaaaagaat ggaatcaaag tgaacttcaa gacccgccac aacattgaag
     6361 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     6421 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     6481 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
     6541 tggatgaact gtacaattaa tactagtagc ggccgctgca ggagtcacta agggttagtt
     6601 agttagatta gcagaaagtc aaaagcctcc gaccggaggc ttttgactaa aacttccctt
     6661 ggggttatca ttggggctca ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc
     6721 gctaaggatg atttctgcta gagatggaat agactggatg gaggcggata aagttgcagg
     6781 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6841 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6901 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6961 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7021 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7141 cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     7201 cgaaaaaacc gccttgcagg gcggttttcg aaggttctct gagctaccaa ctctttgaac
     7261 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     7321 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     7381 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7441 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     7501 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     7561 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     7621 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     7681 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     7741 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     7801 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     7861 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttc
//
