LOCUS       sequence-119835-        3219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Transcriptional Unit: FACS Standard Color Controls - High GFP 
            expression cassette - Kanamycin plasmid 
            [E:J23102:B:BCD2:C:E0040m:D:B0015:F].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3219)
  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 3219)
  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_EF
FEATURES             Location/Qualifiers
     source          1..3219
                     /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..845
                     /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(162..190)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     785..805
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      868..939
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      955..982
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1001..1021
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1022..1079
                     /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(1103..1120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1274..1862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1354..1373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2095..2910)
                     /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(2095..2910)
                     /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     2818..2837
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2902..2921
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3115..3133
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3137..3180)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3183..3204
                     /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 gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      181 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
      241 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
      301 ggccaacact tgtcactact ttcggttatg gtgttcaatg ctttgcgaga tacccagatc
      361 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
      421 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
      481 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
      541 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
      601 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
      661 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
      721 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagatc
      781 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
      841 acaaataata aaggtaggtc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      901 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
      961 ccttcgggtg ggcctttctg cgtttatacg ctatgtcttc tactagtagc ggccgctgca
     1021 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
     1081 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1141 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1201 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1261 cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1681 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1861 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1921 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1981 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgagtc
     2041 ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa
     2101 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2161 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2221 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2281 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     2341 cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt
     2401 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     2461 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     2521 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     2581 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     2641 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     2701 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     2761 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     2821 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctgga
     2881 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     2941 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     3001 ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagct
     3061 cgagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3121 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     3181 tggaattcgc ggccgcttct agagactagt ggaagacat
//
