LOCUS       sequence-192739-        2935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 25.
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: YCe1721 HC_Kan_RFP-p25
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    63..83
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(90..117)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(133..204)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(249..926)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             933..944
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    962..978
                     /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(986..992,993..1010,1011..1016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1031..1052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    1153..1174
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1258..1289
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1290..1392
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1393..2208
                     /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(1466..1485)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2229..2256
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2268..2855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2348..2367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2877..2906)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gagtctgagg ctcgtcctga atgatatcaa gcttgaattc gtttcgtctc gtcgtagaga
       61 ccctgcagcg gccgctacta gtatataaac gcagaaaggc ccacccgaag gtgagccagt
      121 gtgactctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
      181 cgactgagcc tttcgtttta tttgatgcct ggctctagta gcgatctaca ctagcactat
      241 cagcgttatt aagcaccggt ggagtgacga ccttcagcac gttcgtactg ttcaacgatg
      301 gtgtagtctt cgttgtggga ggtgatgtcc agtttgatgt cggttttgta agcacccggc
      361 agctgaaccg gttttttagc catgtaggtg gttttaactt cagcgtcgta gtgaccaccg
      421 tctttcagtt tcagacgcat tttgatttca cctttcagag caccgtcttc cgggtacata
      481 cgttcggtgg aagcttccca acccatggtt tttttctgca taaccggacc gtcggacggg
      541 aagttggtac cacgcagttt aactttgtag atgaactcac cgtcttgcag ggaggagtcc
      601 tgggtaacgg taacaacacc accgtcttcg aagttcataa cacgttccca tttgaaacct
      661 tccgggaagg acagtttcag gtagtccggg atgtcagccg ggtgtttaac gtaagctttg
      721 gaaccgtact ggaactgcgg ggacaggatg tcccaagcga acggcagcgg accacctttg
      781 gtaactttca gtttagcggt ctgggtacct tcgtacggac gaccttcacc ttcaccttcg
      841 atttcgaact cgtgaccgtt aacggaacct tccatacgaa ctttgaaacg catgaactct
      901 ttgataacgt cttcggagga agccatctag tatttctcct ctttctctag tatgtgtgaa
      961 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1021 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1081 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1141 gtttgcgtat tgctctagaa gcggccgcga attcggtctc tcagtcgaga cgtgacgaat
     1201 tctctagata tcgctcaata ctgaccattt aaatcatacc tgacctccat agcagaaagt
     1261 caaaagcctc cgaccggagg cttttgactt gatcggcacg taagaggttc caactttcac
     1321 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1381 aaggaagcta aaatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat
     1441 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1501 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1561 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcaggctaaa ctggctgacg
     1621 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1681 ctcaccactg cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca
     1741 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1801 tgtaattgtc cttttaacgg cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1861 aataacggtt tggttggtgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1921 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     1981 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2041 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2101 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2161 gatatgaata aattgcagtt tcacttgatg ctcgatgagt ttttctgagg gcccaaatgt
     2221 aatcacctgg ctcaccttcg ggtgggcctt tctgcgttgc tggcgttttt ccataggctc
     2281 cgcccccctg acgagcatca caaaaatcga tgctcaagtc agaggtggcg aaacccgaca
     2341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctc ggaaaaagag
     2761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatt ttctaccgaa
     2881 gaaaggccca cccgtgaagg tgagccagtg agttgattgc agtccagtta cgctg
//
