LOCUS       sequence-205826-        6622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBW100ParsR carrying 30hrpR-32hrpS-t-hrpL-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6622)
  AUTHORS   Wang B, Barahona M, Buck M
  TITLE     Engineering modular and tunable genetic amplifiers for scaling 
            transcriptional signals in cascaded gene networks.
  JOURNAL   Nucleic Acids Res. 2014 Aug;42(14):9484-92. doi: 10.1093/nar/gku593.
            Epub 2014 Jul 16.
  PUBMED    25030903
REFERENCE   2  (bases 1 to 6622)
  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: 
            "10.1093/nar/gku593"; journalName: "Nucleic Acids Res"; date: 
            "2014-08"; volume: "42"; issue: "14"; pages: "9484-92"
COMMENT     Sequence Label: pBW102ParsR-Amp32C
FEATURES             Location/Qualifiers
     source          1..6622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..44)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    47..68
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        69..103
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      547..618
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      634..661
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1709..1720
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      2719..2790
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2806..2833
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3067..3078
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3085..3798
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3115..3143)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3738..3758
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3821..3892
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3908..3935
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3942..3962
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3963..4020
                     /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."
     CDS             complement(4178..4993)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4178..4993)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4901..4920
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4985..5004
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5588..6133)
                     /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(5668..5687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6601..6619
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
       61 ttctagagtt tacagctagc tcagtccttg gtattatgct agctcctagg gtcacacagg
      121 aaagtaccat atgtcatttc tgttacccat ccaattgttc aaaattcttg ctgatgaaac
      181 ccgtctgggc atcgttttac tgctcagcga actgggagag ttatgcgtct gcgatctctg
      241 cactgctctc gaccagtcgc agcccaagat ctcccgccac ctggcattgc tgcgtgaaag
      301 cgggctattg ctggaccgca agcaaggtaa gtgggttcat taccgcttat caccgcatat
      361 tccagcatgg gcggcgaaaa ttattgatga ggcctggcga tgtgaacagg aaaaggttca
      421 ggcgattgtc cgcaacctgg ctcgacaaaa ctgttccggg gacagtaaga acatttgcag
      481 ttaaaaattt agctaaacac acatgaattt tcagatgtgt tttatccggg tactagagcc
      541 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      601 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
      661 gtttatatac tagagccaac tcaaaattca cacctattac cttcctctgc acttacacat
      721 tcgttaagtc atatatgttt ttgacttatc cgcttcgaag agagacacta cctgcaatac
      781 tagagtcaca caggaaagta ctagatgagt acaggcatcg ataaggacgt ccgagagtgt
      841 tggggcgtaa ctgcattatc agcgggtcat caaattgcaa tgaatagcgc gtttctggat
      901 atggacttgc tgttgtgcgg ggaaaccggc accggcaagg acacactggc caaccgcatt
      961 cacgagttgt ccagcaggtc gggacccttt gtgggcatga actgcgccgc cattcccgag
     1021 tcgctggcag agagccagtt attcggtgtg gtcaacggtg cattcaccgg cgtatgccgg
     1081 gctcgcgagg gctacataga ggcctccagt ggtggcacct tgtacctgga tgaaatcgac
     1141 agcatgccgt tgagcctgca agccaaactg ctgcgtgtgt tggagagtcg aggtatcgag
     1201 cgtctgggct cgaccgaatt tatcccggtg gatctgcgga tcattgcctc ggcccagcgg
     1261 ccactggatg aactggtgga acaaggactt ttccgtcgcg acctgttttt tcggctcaac
     1321 gtgctgacgc ttcacttgcc agccttgcgc aaacgtcgtg aacagatcct gccattgttc
     1381 gaccagttca cccagggtat cgctgccgag ttcggacgtc ccgctcctgc gctggacagc
     1441 gggcgtgtgc agctgctgct cagccacgac tggccgggca acatccgcga attgaagtct
     1501 gcggccaagc gcttcgtact cggcttcccc ttgctgggcg ccgaccctgt ggaagcgctt
     1561 gaccctgcca cggggctgcg cacgcaaatg cgcatcatcg agaaaatgct catccaggat
     1621 gccttgaagc ggcacaggca caatttcgac gcggtgcttc aggagttgga gttgccaaga
     1681 cgcaccctgt atcaccgcat gaaggaactg ggagttgcag cgccgatcgc tgcgacggcc
     1741 ggggtctaat aaaacgcagg tgtggttatg ctactagagt cacacaggaa agtactagat
     1801 gagtcttgat gaaaggtttg aggatgatct ggacgaggag cgggttccga atctggggat
     1861 agttgccgaa agtatttcgc aactgggtat cgacgtgctg ctatcgggtg agaccggcac
     1921 gggcaaagac acgattgccc gacggattca tgagatgtca ggccgcaaag ggcgcctggt
     1981 ggcgatgaat tgcgcggcca ttccggagtc cctcgccgag agcgagttat tcggcgtggt
     2041 cagcggtgcc tacaccggcg ctgatcgctc cagagtcggt tatgtcgaag cggcgcaggg
     2101 cggcacgctg tacctggatg agatcgatag catgccgctg agcctgcaag ccaaattgct
     2161 gagggtgctg gaaacccgag cgcttgaacg gctgggttcg acgtcgacga tcaagctgga
     2221 tatctgcgtg atcgcctccg cccaatgctc gctggacgac gccgtcgagc gggggcagtt
     2281 tcgtcgcgat ctgtattttc gcctgaacgt cctgacactc aagcttcctc cgctacgtaa
     2341 ccagtctgat cgcatagttc ccctgttcac acgttttacg gccgccgccg cgagggagct
     2401 cggtgttccc gttcccgatg tttgcccact gctgcacaaa gtgctgctgg gccacgactg
     2461 gcccggcaat atccgtgagc tcaaggcggc agccaaacgc catgtgctgg gtttcccctt
     2521 gctgggcgcc gagccgcagg gcgaagagca cttggcctgt gggctcaaat cgcaattgcg
     2581 agtgatcgaa aaagccctga ttcaggagtc gctcaagcgc cacgacaatt gtgtggattc
     2641 ggtaagcctg gaactggacg tgccacgccg tacgctctat cgacgcatca aagaattgca
     2701 gatctaataa ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2761 tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     2821 gggcctttct gcgtttatat actagaggcc ggattatgtc cgctgagtgg gtcacggtcc
     2881 cggatcagtt cccttgcgaa gctgaccgat gtttttgtgc caaaagctgt tgtggcaaaa
     2941 aacggtttgc gcaaagtttt gtattacaaa gaatttcaca ttttaaaata tctttataaa
     3001 tcaatcagtt atttctattt ttaagctggc atggttatcg ctatagggct tgtactacta
     3061 gagattaaag aggagaaata ctagatgcgt aaaggagaag aacttttcac tggagttgtc
     3121 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     3181 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     3241 ctacctgttc catggccaac acttgtcact actttcggtt atggtgttca atgctttgcg
     3301 agatacccag atcatatgaa acagcatgac tttttcaaga gtgccatgcc cgaaggttat
     3361 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     3421 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
     3481 gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc
     3541 atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca caacattgaa
     3601 gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     3661 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     3721 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     3781 atggatgaac tatacaaata ataatactag agccaggcat caaataaaac gaaaggctca
     3841 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     3901 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagtag cggccgctgc
     3961 agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga
     4021 ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4081 cggcgagcgg tatcagctca ctcaaaggcg gtaatctcga gtcccgtcaa gtcagcgtaa
     4141 tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca
     4201 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     4261 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     4321 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     4381 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     4441 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     4501 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     4561 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     4621 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     4681 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     4741 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     4801 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     4861 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     4921 catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt
     4981 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     5041 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
     5101 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca
     5161 acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca
     5221 aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct
     5281 ggtatgagtc agcaacacct tcttcacgag gcagacctca gcgctagcgg agtgtatact
     5341 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     5401 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     5461 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     5521 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     5581 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     5641 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     5701 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     5761 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     5821 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     5881 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     5941 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     6001 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     6061 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     6121 aaaacgatct caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc
     6181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6541 tattccatgg tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6601 aataggcgta tcacgaggca ga
//
