LOCUS       sequence-68365-e        3031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BIOFAB RFP reporter plasmid for measuring promoter 1 + BCD1 
            efficiency..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3031)
  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 3031)
  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: pFAB3665
FEATURES             Location/Qualifiers
     source          1..3031
                     /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             1468..2145
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2178..2249
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2265..2292
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2335..2352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2469..3014)
                     /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(2549..2568)
                     /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 ttaaagtcta acctataggt ataatgtgtg
     1381 gagggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca
     1441 tcttaatcat gcacaggaga ctttctaatg gcgagtagcg aagacgttat caaagagttc
     1501 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
     1561 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
     1621 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
     1681 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
     1741 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
     1801 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
     1861 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
     1921 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
     1981 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
     2041 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
     2101 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaaggatc ggttgtcgag
     2161 taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2221 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2281 ggcctttctg cgtttatagg atcctaactc gagcctaggg atatattccg cttcctcgct
     2341 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg
     2401 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
     2461 aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat
     2521 cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc
     2581 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
     2641 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
     2701 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
     2761 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
     2821 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
     2881 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
     2941 tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac
     3001 caaaacgatc tcaagaagat catcttatta a
//
