LOCUS       sequence-238434-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP Reporter plasmid for B-Rex contain imperfect operator sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  AUTHORS   Liu Y, Landick R, Raman S
  TITLE     A Regulatory NADH/NAD+ Redox Biosensor for Bacteria.
  JOURNAL   ACS Synth Biol. 2019 Feb 15;8(2):264-273. doi: 
            10.1021/acssynbio.8b00485. Epub 2019 Jan 23.
  PUBMED    30633862
REFERENCE   2  (bases 1 to 5334)
  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.8b00485"; journalName: "ACS Synth Biol"; date: 
            "2019-02-15- 15"; volume: "8"; issue: "2"; pages: "264-273"
COMMENT     Sequence Label: pROP-IP-GFP
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..12
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(49..77)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     672..692
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      795..881
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(986..1777)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVITEVSTQLSEVVGAIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(2354..3304)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3352..3574
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4066..4160)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(4980..5014)
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     terminator      complement(5107..5136)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     5207..5225
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaagaggaga aaggtaccat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
       61 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
      121 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
      181 gttccatggc caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac
      241 ccagatcata tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
      301 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
      361 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
      421 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
      481 gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga
      541 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
      601 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
      661 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
      721 gatctctaca aataaaagct tgatatcgaa ttcctgcagc ccgggggatc ccatggtacg
      781 cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      841 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg
      901 gggttatcat tggggctcac tcaaaggcgg taatcagata aaaaaaatcc ttagctttcg
      961 ctaaggatga tttctgctag tattattatt tgccgactac cttggtgatc tcgcctttca
     1021 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
     1081 caagataagc ctgtctagct tcaagtatga cgggctgata ttgggccggc aggcgctcca
     1141 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     1201 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     1261 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     1321 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     1381 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     1441 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     1501 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     1561 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     1621 taactagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     1681 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat ggcgccaact acctctgata
     1741 gttgagtcga tacttcggtg atcaccgctt ccctcataat gtttaacttt gttttagggc
     1801 gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccatggcgt
     1861 aacgcgcttg ctgcttggat gcccgaggca tagactgtac cccaaaaaaa cagtcatagc
     1921 aagccatgaa aaccgccact gcaccgttac caccgctgcg ttcggtcaag gttctggacc
     1981 agttgcgtga gcgcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     2041 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     2101 ggctttgttg aataaatcga acttttgctg agttgaagga tcagctctag tagttacatt
     2161 gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa actcgtaaaa gctctgatgt
     2221 atctatcttt tttacaccgt tttcatctgt gcatatggac agttttccct ttgatatgta
     2281 acggtgaaca gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag
     2341 agccataaga acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt
     2401 gtttttgcgt gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca
     2461 aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt
     2521 ttcttagtcc gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat
     2581 tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa
     2641 cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc
     2701 tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact
     2761 catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta atctctatat
     2821 ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag
     2881 agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact
     2941 ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact
     3001 tggcatagtt tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca
     3061 cagttctcgt catcagctct ctggttgctt tagctaatac accataagca ttttccctac
     3121 tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg
     3181 tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat
     3241 gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag
     3301 acatacatct caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc
     3361 aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct
     3421 ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg
     3481 ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga
     3541 taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt
     3601 acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag
     3661 gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc
     3721 atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct cacggctctg
     3781 gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc aaggaaacta
     3841 cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc gggtctgcta
     3901 tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt tccagtctga
     3961 ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca gtaaggcagc
     4021 ggtatcatca acaggcttac ccgtcttact gtccctagtg cttggattct caccaataaa
     4081 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     4141 tggatctatc aacaggagtc caagcgagct cgtaaacttg gtctgacagc tctagctccg
     4201 gcaaaaaaac gggcaaggtg tcaccaccct gttattttac agctcatcca tgccaccggt
     4261 agaatgacga ccctccgcgc gctcatattg ctctacgatc gtataatctt cattatgaga
     4321 ggtgatgtcc agtttaatat tcacattgta cgcgccaggc agctgcacag gtttcttggc
     4381 tttgtacgtg gttttcactt cagcgtcata atggccgcca tctttcagtt tcaggcgctg
     4441 tttaatttcg cctttcagcg caccatcttc cggatacatg cgttcgctag acgcctccca
     4501 acccatcgtc tttttctgca tcaccgggcc atcagatgga aaattagtac cacgcagttt
     4561 aactttatag atgaactcgc catcctgcag ggaggagtcc tgagtgacgg tcacgacacc
     4621 accatcttca aaattcatta cgcgttccca cttgaaacct tccggaaaag acagcttcag
     4681 atagtccggg atatccgctg ggtgtttaac atacgcttta gaaccgtaca taaattgcgg
     4741 gctcaggatg tcccacgcaa aaggcagcgg gccgccttta gtcactttca gtttggcggt
     4801 ctgggtgcct tcatacggac ggccctcgcc ttcgccttcg atctcgaact cgtgaccgtt
     4861 aacagaaccc tccatgtgaa ctttgaagcg catgaactct ttaatgatag ccatgttatc
     4921 ctcctcgccc ttggaaacca ttcttttcct tcttatcctc gcttaccttc ttgctccatg
     4981 ctagcataat acctagggct gagctagccg taaaggcgcg cccccttttt ctttaaaacc
     5041 gaaaagatta cttcgcgttt gccacctgac gtctaagaaa aggaatattc agcaatttgc
     5101 ccgtgccgaa gaaaggccca cccgtgaagg tgagccagtg agttgattgc tacgtaacaa
     5161 ttccgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5221 gaggcccttt tttcgtcttc acctcgagat gctagcaaaa aaagagtatt tacagctagc
     5281 tcagtcctag gtattattac agccatgtga aatattgagc aaaaagaatt catt
//
