LOCUS       sequence-192650-        2930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 8b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2930)
  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 2930)
  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: YCe1717 HC_Kan_RFP-p8b
FEATURES             Location/Qualifiers
     source          1..2930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    20..41
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    142..163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(178..183,184..201,202..208)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    216..232
                     /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)."
     RBS             250..261
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             268..945
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      990..1061
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1077..1104
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1111..1131
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1210..1241
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1242..1344
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1345..2160
                     /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(1418..1437)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2181..2208
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2220..2807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2300..2319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2829..2858)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 acgtctcgat ccagagaccg aattcgcggc cgcttctaga gcaatacgca aaccgcctct
       61 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      121 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      181 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      241 atactagaga aagaggagaa atactagatg gcttcctccg aagacgttat caaagagttc
      301 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
      361 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
      421 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
      481 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
      541 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
      601 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
      661 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
      721 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
      781 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
      841 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
      901 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg ctgatagtgc
      961 tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1021 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     1081 caccttcggg tgggcctttc tgcgtttata tactagtagc ggccgctgca gggtctctgc
     1141 gtcgagacga attctctaga tatcgctcaa tactgaccat ttaaatcata cctgacctcc
     1201 atagcagaaa gtcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt
     1261 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     1321 ttttcaggag ctaaggaagc taaaatgagc catattcaac gggaaacgtc ttgctcgagg
     1381 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1441 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     1501 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcaggcta
     1561 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1621 gatgcatggt tactcaccac tgcgatccca gggaaaacag cattccaggt attagaagaa
     1681 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1741 tcgattcctg tttgtaattg tccttttaac ggcgatcgcg tatttcgtct cgctcaggcg
     1801 caatcacgaa tgaataacgg tttggttggt gcgagtgatt ttgatgacga gcgtaatggc
     1861 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
     1921 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1981 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2041 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2101 attgataatc ctgatatgaa taaattgcag tttcacttga tgctcgatga gtttttctga
     2161 gggcccaaat gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt
     2221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg
     2281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2641 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2821 ttttctaccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gcagtccagt
     2881 tacgctggag tctgaggctc gtcctgaatg atatcaagct tgaattcgtt
//
