LOCUS       sequence-119876-        4033 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 4033)
  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 4033)
  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..4033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..44
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(263..367)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     435..453
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(457..500)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    503..524
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        543..577
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             652..660
                     /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             670..1383
                     /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
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDYMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(700..728)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1323..1343
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1402..1473
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1489..1516
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1527..1561
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             1636..1644
                     /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             1654..2331
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2347..2418
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2434..2461
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2479..2499
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(2505..2522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2676..3264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2756..2775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3435..4033,1..193,194..262))
                     /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"
ORIGIN
        1 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
       61 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
      181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
      241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
      301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      361 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
      421 cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag
      481 ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agagactagt gggtctcagg
      541 agttgacagc tagctcagtc ctaggtactg tgctagctac tgggcccaag ttcacttaaa
      601 aaggagatca acaatgaaag caattttcgt actgaaacat cttaatcatg ctaaggaggt
      661 tttctaatga tgcgtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      721 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      781 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
      841 ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata cccagatcat
      901 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
      961 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     1021 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     1081 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     1141 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     1201 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1261 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagactac
     1321 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     1381 aaataataaa ggtccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1441 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     1501 ggtgggcctt tctgcgttta tagcttttga cagctagctc agtcctaggt actgtgctag
     1561 ctactgggcc caagttcact taaaaaggag atcaacaatg aaagcaattt tcgtactgaa
     1621 acatcttaat catgctaagg aggttttcta atgatggctt cctccgagga tgttatcaaa
     1681 gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc
     1741 gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc
     1801 aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc
     1861 aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa
     1921 ggtttcaaat gggaacgtgt tatgaacttc gaagatggtg gtgttgttac cgttacccag
     1981 gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc
     2041 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
     2101 atgtacccgg aggatggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
     2161 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
     2221 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga ggactacacc
     2281 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataaaggtcc
     2341 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2401 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     2461 gtttatacgc tagagaccta ctagtagcgg ccgctgcagg cttcctcgct cactgactcg
     2521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac
     2701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3301 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3361 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3421 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3481 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3541 ttaccatctg gccccagtgc tgcaatgata ccgcgcgacc cacgctcacc ggctccagat
     3601 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3661 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3721 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3781 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3841 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3901 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3961 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4021 cggcgaccga gtt
//
