LOCUS       sequence-406247-        6626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBW100ParsR carrying 30hrpR-30hrpS-t-hrpL-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6626)
  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 6626)
  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: pBW103ParsR-Amp30C
FEATURES             Location/Qualifiers
     source          1..6626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..29
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(33..76)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    79..100
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        101..135
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      579..650
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      666..693
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             821..832
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1743..1754
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             1817..1828
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2755..2826
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2842..2869
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3103..3114
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3121..3834
                     /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(3151..3179)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3774..3794
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3857..3928
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3944..3971
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3978..3998
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3999..4056
                     /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(4214..5029)
                     /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(4214..5029)
                     /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     4937..4956
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5021..5040
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5624..6169)
                     /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(5704..5723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
       61 cgctaaggat gatttctgga attcgcggcc gcttctagag tttacagcta gctcagtcct
      121 tggtattatg ctagctccta gggtcacaca ggaaagtacc atatgtcatt tctgttaccc
      181 atccaattgt tcaaaattct tgctgatgaa acccgtctgg gcatcgtttt actgctcagc
      241 gaactgggag agttatgcgt ctgcgatctc tgcactgctc tcgaccagtc gcagcccaag
      301 atctcccgcc acctggcatt gctgcgtgaa agcgggctat tgctggaccg caagcaaggt
      361 aagtgggttc attaccgctt atcaccgcat attccagcat gggcggcgaa aattattgat
      421 gaggcctggc gatgtgaaca ggaaaaggtt caggcgattg tccgcaacct ggctcgacaa
      481 aactgttccg gggacagtaa gaacatttgc agttaaaaat ttagctaaac acacatgaat
      541 tttcagatgt gttttatccg ggtactagag ccaggcatca aataaaacga aaggctcagt
      601 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
      661 acactggctc accttcgggt gggcctttct gcgtttatat actagagcca actcaaaatt
      721 cacacctatt accttcctct gcacttacac attcgttaag tcatatatgt ttttgactta
      781 tccgcttcga agagagacac tacctgcaat actagagatt aaagaggaga aatactagat
      841 gagtacaggc atcgataagg acgtccgaga gtgttggggc gtaactgcat tatcagcggg
      901 tcatcaaatt gcaatgaata gcgcgtttct ggatatggac ttgctgttgt gcggggaaac
      961 cggcaccggc aaggacacac tggccaaccg cattcacgag ttgtccagca ggtcgggacc
     1021 ctttgtgggc atgaactgcg ccgccattcc cgagtcgctg gcagagagcc agttattcgg
     1081 tgtggtcaac ggtgcattca ccggcgtatg ccgggctcgc gagggctaca tagaggcctc
     1141 cagtggtggc accttgtacc tggatgaaat cgacagcatg ccgttgagcc tgcaagccaa
     1201 actgctgcgt gtgttggaga gtcgaggtat cgagcgtctg ggctcgaccg aatttatccc
     1261 ggtggatctg cggatcattg cctcggccca gcggccactg gatgaactgg tggaacaagg
     1321 acttttccgt cgcgacctgt tttttcggct caacgtgctg acgcttcact tgccagcctt
     1381 gcgcaaacgt cgtgaacaga tcctgccatt gttcgaccag ttcacccagg gtatcgctgc
     1441 cgagttcgga cgtcccgctc ctgcgctgga cagcgggcgt gtgcagctgc tgctcagcca
     1501 cgactggccg ggcaacatcc gcgaattgaa gtctgcggcc aagcgcttcg tactcggctt
     1561 ccccttgctg ggcgccgacc ctgtggaagc gcttgaccct gccacggggc tgcgcacgca
     1621 aatgcgcatc atcgagaaaa tgctcatcca ggatgccttg aagcggcaca ggcacaattt
     1681 cgacgcggtg cttcaggagt tggagttgcc aagacgcacc ctgtatcacc gcatgaagga
     1741 actgggagtt gcagcgccga tcgctgcgac ggccggggtc taataaaacg caggtgtggt
     1801 tatgctacta gagattaaag aggagaaata ctagatgagt cttgatgaaa ggtttgagga
     1861 tgatctggac gaggagcggg ttccgaatct ggggatagtt gccgaaagta tttcgcaact
     1921 gggtatcgac gtgctgctat cgggtgagac cggcacgggc aaagacacga ttgcccgacg
     1981 gattcatgag atgtcaggcc gcaaagggcg cctggtggcg atgaattgcg cggccattcc
     2041 ggagtccctc gccgagagcg agttattcgg cgtggtcagc ggtgcctaca ccggcgctga
     2101 tcgctccaga gtcggttatg tcgaagcggc gcagggcggc acgctgtacc tggatgagat
     2161 cgatagcatg ccgctgagcc tgcaagccaa attgctgagg gtgctggaaa cccgagcgct
     2221 tgaacggctg ggttcgacgt cgacgatcaa gctggatatc tgcgtgatcg cctccgccca
     2281 atgctcgctg gacgacgccg tcgagcgggg gcagtttcgt cgcgatctgt attttcgcct
     2341 gaacgtcctg acactcaagc ttcctccgct acgtaaccag tctgatcgca tagttcccct
     2401 gttcacacgt tttacggccg ccgccgcgag ggagctcggt gttcccgttc ccgatgtttg
     2461 cccactgctg cacaaagtgc tgctgggcca cgactggccc ggcaatatcc gtgagctcaa
     2521 ggcggcagcc aaacgccatg tgctgggttt ccccttgctg ggcgccgagc cgcagggcga
     2581 agagcacttg gcctgtgggc tcaaatcgca attgcgagtg atcgaaaaag ccctgattca
     2641 ggagtcgctc aagcgccacg acaattgtgt ggattcggta agcctggaac tggacgtgcc
     2701 acgccgtacg ctctatcgac gcatcaaaga attgcagatc taataaccag gcatcaaata
     2761 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2821 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacta
     2881 gaggccggat tatgtccgct gagtgggtca cggtcccgga tcagttccct tgcgaagctg
     2941 accgatgttt ttgtgccaaa agctgttgtg gcaaaaaacg gtttgcgcaa agttttgtat
     3001 tacaaagaat ttcacatttt aaaatatctt tataaatcaa tcagttattt ctatttttaa
     3061 gctggcatgg ttatcgctat agggcttgta ctactagaga ttaaagagga gaaatactag
     3121 atgcgtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     3181 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
     3241 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     3301 gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca tatgaaacag
     3361 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     3421 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     3481 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
     3541 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     3601 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     3661 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     3721 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     3781 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaataataa
     3841 tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3901 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     3961 gcctttctgc gtttatatac tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg
     4021 tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc
     4081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4141 aaggcggtaa tctcgagtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat
     4201 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     4261 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     4321 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     4381 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     4441 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     4501 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     4561 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     4621 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     4681 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     4741 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     4801 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     4861 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     4921 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     4981 aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg
     5041 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     5101 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     5161 ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc
     5221 aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc
     5281 tcactttctg gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt
     5341 cacgaggcag acctcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
     5401 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     5461 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     5521 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     5581 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     5641 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     5701 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     5761 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     5821 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     5881 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     5941 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     6001 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     6061 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     6121 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     6181 tattaagggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6541 aagtggtcct gcaactttat ccgcctccat ccagtctatt ccatggtgcc acctgacgtc
     6601 taagaaacca ttattatcat gacatt
//
