LOCUS       sequence-114975-        3176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Transcriptional Unit: FACS Standard Color Controls - High RFP 
            expression cassette - Kanamycin plasmid 
            [E:J23102:B:BCD2:C:E1010m:D:B0015:F].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3176)
  AUTHORS   Iverson SV, Haddock TL, Beal J, Densmore DM
  TITLE     CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part 
            Library Enable Rapid Combinatorial Design for Synthetic and 
            Traditional Biology.
  JOURNAL   ACS Synth Biol. 2016 Jan 15;5(1):99-103. doi: 
            10.1021/acssynbio.5b00124. Epub 2015 Nov 4.
  PUBMED    26479688
REFERENCE   2  (bases 1 to 3176)
  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.5b00124"; journalName: "ACS Synth Biol"; date: 
            "2016-01-15- 15"; volume: "5"; issue: "1"; pages: "99-103"
COMMENT     Sequence Label: pJ02B2Rm_EF
FEATURES             Location/Qualifiers
     source          1..3176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             114..122
                     /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             132..809
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      825..896
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      912..939
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    958..978
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      979..1036
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1060..1077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1231..1819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1311..1330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2052..2867)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2052..2867)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2775..2794
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2859..2878
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3072..3090
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3094..3137)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3140..3161
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gcttttgaca gctagctcag tcctaggtac tgtgctagct actgggccca agttcactta
       61 aaaaggagat caacaatgaa agcaattttc gtactgaaac atcttaatca tgctaaggag
      121 gttttctaat gatggcttcc tccgaggatg ttatcaaaga gttcatgcgt ttcaaagttc
      181 gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc
      241 cgtacgaagg tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg
      301 cttgggacat cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg
      361 ctgacatccc ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta
      421 tgaacttcga agatggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg
      481 agttcatcta caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc
      541 agaaaaaaac catgggttgg gaagcttcca ccgaacgtat gtacccggag gatggtgctc
      601 tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag
      661 ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca
      721 tcaaactgga catcacctcc cacaacgagg actacaccat cgttgaacag tacgaacgtg
      781 ctgaaggtcg tcactccacc ggtgcttaat aaaggtccag gcatcaaata aaacgaaagg
      841 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
      901 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacgcta tgtcttctac
      961 tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc
     1021 tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg
     1081 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca
     1261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1981 gtctgacagc tcgagtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta
     2041 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     2101 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     2161 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     2221 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     2281 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     2341 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2401 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     2461 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     2521 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     2581 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     2641 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     2701 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     2761 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2821 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     2881 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     2941 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     3001 gagttgaagg atcagctcga gtgccacctg acgtctaaga aaccattatt atcatgacat
     3061 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     3121 tcgctaagga tgatttctgg aattcgcggc cgcttctaga gactagtgga agacat
//
