LOCUS       sequence-192707-        2935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 14.
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: YCe1709 HC_Kan_RFP-p14
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    52..72
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(79..106)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(122..193)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(238..915)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             922..933
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    951..967
                     /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(975..981,982..999,1000..1005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1020..1041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    1142..1163
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1247..1278
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1279..1381
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1382..2197
                     /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(1455..1474)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2218..2245
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2257..2844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2337..2356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2866..2895)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tcgtcctgaa tgatatcaag cttgaattcg tttcgtctcg ttgcagagac cctgcagcgg
       61 ccgctactag tatataaacg cagaaaggcc cacccgaagg tgagccagtg tgactctagt
      121 agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
      181 ttcgttttat ttgatgcctg gctctagtag cgatctacac tagcactatc agcgttatta
      241 agcaccggtg gagtgacgac cttcagcacg ttcgtactgt tcaacgatgg tgtagtcttc
      301 gttgtgggag gtgatgtcca gtttgatgtc ggttttgtaa gcacccggca gctgaaccgg
      361 ttttttagcc atgtaggtgg ttttaacttc agcgtcgtag tgaccaccgt ctttcagttt
      421 cagacgcatt ttgatttcac ctttcagagc accgtcttcc gggtacatac gttcggtgga
      481 agcttcccaa cccatggttt ttttctgcat aaccggaccg tcggacggga agttggtacc
      541 acgcagttta actttgtaga tgaactcacc gtcttgcagg gaggagtcct gggtaacggt
      601 aacaacacca ccgtcttcga agttcataac acgttcccat ttgaaacctt ccgggaagga
      661 cagtttcagg tagtccggga tgtcagccgg gtgtttaacg taagctttgg aaccgtactg
      721 gaactgcggg gacaggatgt cccaagcgaa cggcagcgga ccacctttgg taactttcag
      781 tttagcggtc tgggtacctt cgtacggacg accttcacct tcaccttcga tttcgaactc
      841 gtgaccgtta acggaacctt ccatacgaac tttgaaacgc atgaactctt tgataacgtc
      901 ttcggaggaa gccatctagt atttctcctc tttctctagt atgtgtgaaa ttgttatccg
      961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1141 gctctagaag cggccgcgaa ttcggtctct gtagcgagac gtgacgaatt ctctagatat
     1201 cgctcaatac tgaccattta aatcatacct gacctccata gcagaaagtc aaaagcctcc
     1261 gaccggaggc ttttgacttg atcggcacgt aagaggttcc aactttcacc ataatgaaat
     1321 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1381 aatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga
     1441 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1501 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1561 cgttgccaat gatgttacag atgagatggt caggctaaac tggctgacgg aatttatgcc
     1621 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1681 gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1741 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1801 ttttaacggc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     1861 ggttggtgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1921 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1981 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2041 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2101 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2161 attgcagttt cacttgatgc tcgatgagtt tttctgaggg cccaaatgta atcacctggc
     2221 tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc cataggctcc gcccccctga
     2281 cgagcatcac aaaaatcgat gctcaagtca gaggtggcga aacccgacag gactataaag
     2341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2701 agtatttggt atctgcgctc tgctgaagcc agttacctcg gaaaaagagt tggtagctct
     2761 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2821 acgcgcagaa aaaaaggatc tcaagaagat cctttgattt tctaccgaag aaaggcccac
     2881 ccgtgaaggt gagccagtga gttgattgca gtccagttac gctggagtct gaggc
//
