LOCUS       sequence-192649-        2935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 8a.
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: YCe1730 HC_Kan_RFP-p8a
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    20..40
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(47..74)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(90..161)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(206..883)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             890..901
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    919..935
                     /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(943..949,950..967,968..973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    988..1009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    1110..1131
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1215..1246
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1247..1349
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1350..2165
                     /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(1423..1442)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2186..2213
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2225..2812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2305..2324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2834..2863)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tcgtctcggg atagagaccc tgcagcggcc gctactagta tataaacgca gaaaggccca
       61 cccgaaggtg agccagtgtg actctagtag agagcgttca ccgacaaaca acagataaaa
      121 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc tctagtagcg
      181 atctacacta gcactatcag cgttattaag caccggtgga gtgacgacct tcagcacgtt
      241 cgtactgttc aacgatggtg tagtcttcgt tgtgggaggt gatgtccagt ttgatgtcgg
      301 ttttgtaagc acccggcagc tgaaccggtt ttttagccat gtaggtggtt ttaacttcag
      361 cgtcgtagtg accaccgtct ttcagtttca gacgcatttt gatttcacct ttcagagcac
      421 cgtcttccgg gtacatacgt tcggtggaag cttcccaacc catggttttt ttctgcataa
      481 ccggaccgtc ggacgggaag ttggtaccac gcagtttaac tttgtagatg aactcaccgt
      541 cttgcaggga ggagtcctgg gtaacggtaa caacaccacc gtcttcgaag ttcataacac
      601 gttcccattt gaaaccttcc gggaaggaca gtttcaggta gtccgggatg tcagccgggt
      661 gtttaacgta agctttggaa ccgtactgga actgcgggga caggatgtcc caagcgaacg
      721 gcagcggacc acctttggta actttcagtt tagcggtctg ggtaccttcg tacggacgac
      781 cttcaccttc accttcgatt tcgaactcgt gaccgttaac ggaaccttcc atacgaactt
      841 tgaaacgcat gaactctttg ataacgtctt cggaggaagc catctagtat ttctcctctt
      901 tctctagtat gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1081 acgcgcgggg agaggcggtt tgcgtattgc tctagaagcg gccgcgaatt cggtctctgc
     1141 ctcgagacgt gacgaattct ctagatatcg ctcaatactg accatttaaa tcatacctga
     1201 cctccatagc agaaagtcaa aagcctccga ccggaggctt ttgacttgat cggcacgtaa
     1261 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     1321 cgagattttc aggagctaag gaagctaaaa tgagccatat tcaacgggaa acgtcttgct
     1381 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1441 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     1501 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1561 ggctaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1621 ctgatgatgc atggttactc accactgcga tcccagggaa aacagcattc caggtattag
     1681 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     1741 tgcattcgat tcctgtttgt aattgtcctt ttaacggcga tcgcgtattt cgtctcgctc
     1801 aggcgcaatc acgaatgaat aacggtttgg ttggtgcgag tgattttgat gacgagcgta
     1861 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     1921 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     1981 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2041 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2101 atggtattga taatcctgat atgaataaat tgcagtttca cttgatgctc gatgagtttt
     2161 tctgagggcc caaatgtaat cacctggctc accttcgggt gggcctttct gcgttgctgg
     2221 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgatgc tcaagtcaga
     2281 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2341 tgcgctctcc tgttccgacc ctgtcgctta ccggatacct gtccgccttt ctcccttcgg
     2401 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2461 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2521 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2581 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2641 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2701 ttacctcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2761 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2821 tttgattttc taccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgcagt
     2881 ccagttacgc tggagtctga ggctcgtcct gaatgatatc aagcttgaat tcgtt
//
