LOCUS       sequence-447362-        5185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23117_RC2_mTagBFP_Buj_mRFP dual operon reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5185)
  AUTHORS   Cardiff RAL, Faulkner ID, Beall JG, Carothers JM, Zalatan JG
  TITLE     
  JOURNAL   Nucleic Acids Res. 2024 May 22;52(9):5406-5419. doi: 
            10.1093/nar/gkae275.
  PUBMED    38613390
REFERENCE   2  (bases 1 to 5185)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2024-05-22- 22"; volume: "52"; issue: 
            "9"; pages: "5406-5419"
COMMENT     Sequence Label: pRC241
FEATURES             Location/Qualifiers
     source          1..5185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..217
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             250..258
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             264..965
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     RBS             981..992
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             999..1676
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1709..1780
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1796..1823
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2354..3304)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTMKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(4066..4160)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(4190..4981,4982..5050))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4813..4832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5051..5155)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aattccgacg tcagcatttg cgatcattca cgcagcgctt attcagttgc tcactgcgat
       61 gtcataatca tcgctacgag ctgtgaaaga tgcataaagc tcgtacgacg cgttcgctcg
      121 tctcctcact tctcctacgg agcgttctgg acacaacgtc gtcttgaagt tgcgattata
      181 gattgacagc tagctcagtc ctagggattg tgctagcgtt cgaaaaagtt ctcacgaggg
      241 agcacagagt aaggaggttt tatatgagcg agctgattaa ggagaacatg cacatgaagc
      301 tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca
      361 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
      421 tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca
      481 cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag
      541 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
      601 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga
      661 tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg
      721 gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa
      781 acgccaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct
      841 actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc
      901 agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta
      961 attaaggatc cgaattcatt aaagaggaga aaggtaccat ggcgagtagc gaagacgtta
     1021 tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg
     1081 aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag
     1141 ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg
     1201 gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc
     1261 cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta
     1321 cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca
     1381 acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg
     1441 aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga
     1501 aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc
     1561 agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact
     1621 acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaaggat
     1681 ccaaactcga gtaaggatct ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1741 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     1801 accttcgggt gggcctttct gcgtttatac ctagggtacg ggttttgctg cccgcaaacg
     1861 ggctgttctg gtgttgctag tttgttatca gaatcgcaga tccggcttca gccggtttgc
     1921 cggctgaaag cgctatttct tccagaattg ccatgatttt ttccccacgg gaggcgtcac
     1981 tggctcccgt gttgtcggca gctttgattc gataagcagc atcgcctgtt tcaggctgtc
     2041 tatgtgtgac tgttgagctg taacaagttg tctcaggtgt tcaatttcat gttctagttg
     2101 ctttgtttta ctggtttcac ctgttctatt aggtgttaca tgctgttcat ctgttacatt
     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 tctttcattg
     3181 tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat
     3241 gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag
     3301 acatacatct caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc
     3361 aatgataatt actagtcctt ttcccgggtg atctgggtat 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 gtctgacagt taccaatgct
     4201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4321 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4381 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4441 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4501 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4561 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4621 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4681 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4741 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4801 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4861 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4921 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4981 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5041 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5101 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5161 ttccccgaaa agtgccacct gtggc
//
