LOCUS       sequence-257698-        5236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Device: FACS Standard Color Controls - High RFP (first unit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5236)
  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 5236)
  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:Gm_AF
FEATURES             Location/Qualifiers
     source          1..5236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             68..76
                     /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             86..763
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      779..850
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      866..893
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        904..938
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             1013..1021
                     /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             1031..1744
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1061..1089)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1684..1704
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1767..1838
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1854..1881
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1899..1919
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1925..1942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2096..2684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2176..2195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2855..3646,3647..3715))
                     /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     3478..3497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3716..3820)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3888..3906
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3910..3953)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3956..3977
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 cccaagttca cttaaaaagg agatcaacaa tgaaagcaat tttcgtactg aaacatctta
       61 atcatgctaa ggaggttttc taatgatggc ttcctccgag gatgttatca aagagttcat
      121 gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga
      181 aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg
      241 tccgctgccg ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta
      301 cgttaaacac ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa
      361 atgggaacgt gttatgaact tcgaagatgg tggtgttgtt accgttaccc aggactcctc
      421 cctgcaagac ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga
      481 cggtccggtt atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc
      541 ggaggatggt gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca
      601 ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc
      661 ttacaaaacc gacatcaaac tggacatcac ctcccacaac gaggactaca ccatcgttga
      721 acagtacgaa cgtgctgaag gtcgtcactc caccggtgct taataaaggt ccaggcatca
      781 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
      841 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag
      901 cttttgacag ctagctcagt cctaggtact gtgctagcta ctgggcccaa gttcacttaa
      961 aaaggagatc aacaatgaaa gcaattttcg tactgaaaca tcttaatcat gctaaggagg
     1021 ttttctaatg atgcgtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     1081 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     1141 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
     1201 gccaacactt gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca
     1261 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     1321 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     1381 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     1441 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
     1501 aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca
     1561 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     1621 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagatca
     1681 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     1741 caaataataa aggtaggtcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1801 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     1861 cttcgggtgg gcctttctgc gtttatacgc tagagaccta ctagtagcgg ccgctgcagg
     1921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2101 acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2521 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2701 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2761 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2821 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2881 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2941 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3001 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3061 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3121 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3181 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3241 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3301 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3361 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3421 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3481 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3541 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3601 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3661 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3721 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3781 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3841 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3901 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt
     3961 cgcggccgct tctagagact agtgggtctc aggagttgac agctagctca gtcctaggga
     4021 aggtgcgaat aagcggggaa attcttctcg gctgactcag tcatttcatt tcttcatgtt
     4081 tgagccgatt ttttctcccg taaatgcctt gaatcagcct atttagaccg tttcttcgcc
     4141 atttaaggcg ttatccccag tttttagtga gatctctccc actgacgtat catttggtcc
     4201 gcccgaaaca ggttggccag cgtgaataac atcgccagtt ggttatcgtt tttcagcaac
     4261 cccttgtatc tggctttcac gaagccgaac tgtcgcttga tgatgcgaaa tgggtgctcc
     4321 accctggccc ggatgctggc tttcatgtat tcgatgttga tggccgtttt gttcttgcgt
     4381 ggatgctgtt tcaaggttct taccttgccg gggcgctcgg cgatcagcca gtccacatcc
     4441 acctcggcca gctcctcgcg ctgtggcgcc ccttggtagc cggcatcggc tgagacaaat
     4501 tgctcctctc catgcagcag attacccagc tgattgaggt catgctcgtt ggccgcggtg
     4561 gtgaccaggc tgtgggtcag gccactcttg gcatcgacac caatgtgggc cttcatgcca
     4621 aagtgccact gattgccttt cttggtctga tgcatctccg gatcgcgttg ctgctctttg
     4681 ttcttggtcg agctgggtgc ctcaatgatg gtggcatcga ccaaggtgcc ttgagtcatc
     4741 atgacgcctg cttcggccag ccagcgattg atggtcttga acaattggcg ggccagttga
     4801 tgctgctcca gcaggtggcg gaaattcatg atggtggtgc ggtccggcaa ggcgctatcc
     4861 agggataacc gggcaaacag acgcatggag gcgatttcgt acagagcatc ttccatcgcg
     4921 ccatcgctca ggttgtacca atgctgcatg cagtgaatgc gtagcatggt ttccagcgga
     4981 taaggtcgcc ggccattacc agccttgggg taaaacggct cgatgacttc caccatgttt
     5041 tgccatggca gaatctgctc catgcgggac aagaaaatct cttttctggt ctgacggcgc
     5101 ttactgctga attcactgtc ggcgaaggta agttgatgac tcatgatgaa ccctgttcta
     5161 tggctccaga tgacaaacat gatctcatat cagggacttg ttcgcacctt ccctaggtac
     5221 tgtgctagct actggg
//
