LOCUS       sequence-257697-        5375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Device: FACS Standard Color Controls - High GFP (first unit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5375)
  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 5375)
  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: pJ02B2Gm:Rm_AF
FEATURES             Location/Qualifiers
     source          1..5375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             32..40
                     /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             50..763
                     /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(80..108)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     703..723
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      786..857
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      873..900
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        911..945
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             1020..1028
                     /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             1038..1715
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1731..1802
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1818..1845
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1863..1883
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1889..1906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2060..2648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2140..2159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2819..3610,3611..3679))
                     /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     3442..3461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3680..3784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3852..3870
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3874..3917)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3920..3941
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 caattttcgt actgaaacat cttaatcatg ctaaggaggt tttctaatga tgcgtaaagg
       61 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
      121 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
      181 taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt
      241 cggttatggt gttcaatgct ttgcgagata cccagatcat atgaaacagc atgacttttt
      301 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      361 gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
      421 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
      481 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa
      541 cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca
      601 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca
      661 atctgccctt tcgaaagatc ccaacgaaaa gagagatcac atggtccttc ttgagtttgt
      721 aacagctgct gggattacac atggcatgga tgaactatac aaataataaa ggtaggtcca
      781 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
      841 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
      901 tttatagctt ttgacagcta gctcagtcct aggtactgtg ctagctactg ggcccaagtt
      961 cacttaaaaa ggagatcaac aatgaaagca attttcgtac tgaaacatct taatcatgct
     1021 aaggaggttt tctaatgatg gcttcctccg aggatgttat caaagagttc atgcgtttca
     1081 aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag
     1141 gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc
     1201 cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac
     1261 acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac
     1321 gtgttatgaa cttcgaagat ggtggtgttg ttaccgttac ccaggactcc tccctgcaag
     1381 acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg
     1441 ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaggatg
     1501 gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg
     1561 ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa
     1621 ccgacatcaa actggacatc acctcccaca acgaggacta caccatcgtt gaacagtacg
     1681 aacgtgctga aggtcgtcac tccaccggtg cttaataaag gtccaggcat caaataaaac
     1741 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1801 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acgctagaga
     1861 cctactagta gcggccgctg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1921 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1981 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2041 aggccgcgtt gctggcgttt ttccacaggc tccgcccccc tgacgagcat cacaaaaatc
     2101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2461 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2581 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2641 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2701 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2761 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2821 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2881 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2941 gtgctgcaat gataccgcgc gacccacgct caccggctcc agatttatca gcaataaacc
     3001 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3061 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3121 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3181 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3241 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3301 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3361 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3421 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3481 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3541 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3601 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3661 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3721 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3781 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3841 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     3901 tcgctaagga tgatttctgg aattcgcggc cgcttctaga gactagtggg tctcaggagt
     3961 tgacagctag ctcagtccta ggtactgtgc tctgagagat cccctcataa tttccccaaa
     4021 gcgtaaccat gtgtgaataa attttgagct agtagggttg cagccacgag taagtcttcc
     4081 cttgttattg tgtagccaga atgccgcaaa acttccatgc ctaagcgaac tgttgagagt
     4141 acgtttcgat ttctgactgt gttagcctgg aagtgcttgt cccaaccttg tttctgagca
     4201 tgaacgcccg caagccaaca tgttagttga agcatcaggg cgattagcag catgatatca
     4261 aaacgctctg agctgctcgt tcggctatgg cgtaggccta gtccgtaggc aggacttttc
     4321 aagtctcgga aggtttcttc aatctgcatt cgcttcgaat agatattaac aagttgtttg
     4381 ggtgttcgaa tttcaacagg taagttagtt gctagaaccc atggctcctt tgccgacgct
     4441 gagtagattt taggtgacgg gtggtgacaa tgagtccgtg tcgagcgctg attttttcgg
     4501 cctttagagc gagatttata caatagaatt tggcatgaga ttggattgct tttagtcagc
     4561 ctcttatagc ctaaagtctt tgagtgacta gatgacatat catgtaagtt gctgataggt
     4621 ttccagtttt ccgctcctag gtctgcatat tgtacttttc ctcttactcg acttaaccag
     4681 taccaaccca gcttctcaac ggatttatac catggcactt taaagccagc atcactgaca
     4741 atgagcggtg tggtgttact cggtagaatg ctcgcaaggt cggctagaaa ttggtcatga
     4801 gctttctttg aacattgctc tgaaagcggg aacgctttct cataaagagt aacagaacga
     4861 ccgtgtagtg cgactgaagc tcgcaatacc ataagtcgtt tttgctcacg aatatcagac
     4921 cagtcaacaa gtacaatggg catcgtattg cccgaacaga taaagctagc atgccaacgg
     4981 tatacagcga gtcgctcttt gtggaggtga cgattaccta acaatcggtc gattcgtttg
     5041 atgttatgtt ttgttctcgc tttggttggc aggttacggc caagttcggt aagagtgaga
     5101 gttttacagt caagtaatgc gtggcaagcc aacgttaagc tgttgagtcg ttttaagtgt
     5161 aattcggggc agaattggta aagagagtcg tgtaaaatat cgagttcgca catcttgttg
     5221 tctgattatt gatttttcgc gaaaccattt gatcatatga caagatgtgt atccacctta
     5281 acttaatgat ttttaccaaa atcattaggg gattcatcag tactgtgcta gctactgggc
     5341 ccaagttcac ttaaaaagga gatcaacaat gaaag
//
