LOCUS       sequence-44856-e        2970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2970)
  AUTHORS   Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, 
            Keasling JD
  TITLE     BglBrick vectors and datasheets: A synthetic biology platform for 
            gene expression.
  JOURNAL   J Biol Eng. 2011 Sep 20;5:12.
  PUBMED    21933410
REFERENCE   2  (bases 1 to 2970)
  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"; journalName: "J Biol 
            Eng."; date: "2011-09-20"; volume: "5"; pages: "12"
COMMENT     Sequence Label: pBbE0k-RFP
FEATURES             Location/Qualifiers
     source          1..2970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             44..55
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             62..739
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      784..855
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      871..898
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      952..1023
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1039..1066
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1224..1812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1304..1323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1900..1994)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2025..2819)
                     /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(2136..2155)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2746..2765
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ttgacagcta gctcagtcct aggtactgtg ctagctacta gtgaaagagg agaaatacta
       61 gatggcttcc tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg
      121 ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg
      181 tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat
      241 cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc
      301 ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga
      361 agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta
      421 caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac
      481 catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga
      541 aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac
      601 ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga
      661 catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg
      721 tcactccacc ggtgcttaat aacgctgata gtgctagtgt agatcgctac tagagccagg
      781 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
      841 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
      901 tatatactag aagcggccgg gatcctaact cgagtaagga tctccaggca tcaaataaaa
      961 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1021 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacctagggc
     1081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1861 aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc
     1921 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     1981 tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca
     2041 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     2101 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     2161 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     2221 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     2281 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     2341 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     2401 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     2461 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     2521 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     2581 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     2641 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     2701 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     2761 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     2821 cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt
     2881 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     2941 ccagctggca attccgaatt catgagatct
//
