LOCUS       sequence-68366-e        3030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BIOFAB RFP reporter plasmid for measuring promoter 2 + BCD1 
            efficiency..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3030)
  AUTHORS   Mutalik VK, Guimaraes JC, Cambray G, Lam C, Christoffersen MJ, Mai 
            QA, Tran AB, Paull M, Keasling JD, Arkin AP, Endy D
  TITLE     Precise and reliable gene expression via standard transcription and 
            translation initiation elements.
  JOURNAL   Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 
            2013 Mar 10.
  PUBMED    23474465
REFERENCE   2  (bases 1 to 3030)
  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: 
            "10.1038/nmeth.2404"; journalName: "Nat Methods"; date: "2013-04"; 
            volume: "10"; issue: "4"; pages: "354-60"
COMMENT     Sequence Label: pFAB3677
FEATURES             Location/Qualifiers
     source          1..3030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(97..191)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(222..1016)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(333..352)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     943..962
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             1467..2144
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2177..2248
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2264..2291
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2334..2351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2468..3013)
                     /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(2548..2567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
       61 cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg
      121 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
      181 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
      241 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
      301 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
      361 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
      421 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
      481 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
      541 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
      601 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
      661 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
      721 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
      781 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
      841 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
      901 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
      961 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     1021 aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca gatccttggc
     1081 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     1141 gctggcaatt ccgacgtcga attcaaaaga tcttaagtaa gtaagagtat acgtatatcg
     1201 gctaataacg tattaaggcg cttcggcgcc tttttttatg ggggtatttt catcccaatc
     1261 cacacgtcca acgcacagca aacaccacgt cgaccctatc agctgcgtgc tttctatgag
     1321 tcgttgctgc ataacaaaaa gagtattgac ttcgcatctt tttgtaccta taatgtgtgg
     1381 agggcccaag ttcacttaaa aaggagatca acaatgaaag caattttcgt actgaaacat
     1441 cttaatcatg cacaggagac tttctaatgg cgagtagcga agacgttatc aaagagttca
     1501 tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg
     1561 aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg
     1621 gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt
     1681 acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca
     1741 aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct
     1801 ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg
     1861 acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc
     1921 cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc
     1981 actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg
     2041 cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg
     2101 aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaaggatcg gttgtcgagt
     2161 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2221 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     2281 gcctttctgc gtttatagga tcctaactcg agcctaggga tatattccgc ttcctcgctc
     2341 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     2401 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     2461 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     2521 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     2581 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     2641 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     2701 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     2761 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     2821 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     2881 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     2941 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3001 aaaacgatct caagaagatc atcttattaa
//
