LOCUS       sequence-192732-        2935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 21.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2935)
  AUTHORS   Martella A, Matjusaitis M, Auxillos J, Pollard SM, Cai Y
  TITLE     EMMA: An Extensible Mammalian Modular Assembly Toolkit for the Rapid
            Design and Production of Diverse Expression Vectors.
  JOURNAL   ACS Synth Biol. 2017 Apr 24. doi: 10.1021/acssynbio.7b00016.
  PUBMED    28418644
REFERENCE   2  (bases 1 to 2935)
  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"; journalName: "ACS Synth 
            Biol. 2017 Apr 24. doi: 10.1021/acssynbio.7b00016."
COMMENT     Sequence Label: YCe1713 HC_Kan_RFP-p21
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    48..68
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(75..102)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(118..189)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(234..911)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             918..929
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    947..963
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(971..977,978..995,996..1001))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1016..1037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    1138..1159
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1243..1274
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1275..1377
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1378..2193
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1451..1470)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2214..2241
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2253..2840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2333..2352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2862..2891)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cctgaatgat atcaagcttg aattcgtttc gtctcgcaac agagaccctg cagcggccgc
       61 tactagtata taaacgcaga aaggcccacc cgaaggtgag ccagtgtgac tctagtagag
      121 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
      181 ttttatttga tgcctggctc tagtagcgat ctacactagc actatcagcg ttattaagca
      241 ccggtggagt gacgaccttc agcacgttcg tactgttcaa cgatggtgta gtcttcgttg
      301 tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac ccggcagctg aaccggtttt
      361 ttagccatgt aggtggtttt aacttcagcg tcgtagtgac caccgtcttt cagtttcaga
      421 cgcattttga tttcaccttt cagagcaccg tcttccgggt acatacgttc ggtggaagct
      481 tcccaaccca tggttttttt ctgcataacc ggaccgtcgg acgggaagtt ggtaccacgc
      541 agtttaactt tgtagatgaa ctcaccgtct tgcagggagg agtcctgggt aacggtaaca
      601 acaccaccgt cttcgaagtt cataacacgt tcccatttga aaccttccgg gaaggacagt
      661 ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag ctttggaacc gtactggaac
      721 tgcggggaca ggatgtccca agcgaacggc agcggaccac ctttggtaac tttcagttta
      781 gcggtctggg taccttcgta cggacgacct tcaccttcac cttcgatttc gaactcgtga
      841 ccgttaacgg aaccttccat acgaactttg aaacgcatga actctttgat aacgtcttcg
      901 gaggaagcca tctagtattt ctcctctttc tctagtatgt gtgaaattgt tatccgctca
      961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgctc
     1141 tagaagcggc cgcgaattcg gtctcttcca cgagacgtga cgaattctct agatatcgct
     1201 caatactgac catttaaatc atacctgacc tccatagcag aaagtcaaaa gcctccgacc
     1261 ggaggctttt gacttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     1321 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     1381 agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct
     1441 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1501 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1561 gccaatgatg ttacagatga gatggtcagg ctaaactggc tgacggaatt tatgcctctt
     1621 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1681 ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1741 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1801 aacggcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1861 ggtgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1921 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1981 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2041 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2101 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2161 cagtttcact tgatgctcga tgagtttttc tgagggccca aatgtaatca cctggctcac
     2221 cttcgggtgg gcctttctgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2281 catcacaaaa atcgatgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2461 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2521 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2581 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2641 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2701 tttggtatct gcgctctgct gaagccagtt acctcggaaa aagagttggt agctcttgat
     2761 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2821 gcagaaaaaa aggatctcaa gaagatcctt tgattttcta ccgaagaaag gcccacccgt
     2881 gaaggtgagc cagtgagttg attgcagtcc agttacgctg gagtctgagg ctcgt
//
