LOCUS       sequence-192632-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2933)
  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 2933)
  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: YCe1500 HC_Kan_RFP-p2
FEATURES             Location/Qualifiers
     source          1..2933
                     /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      1213..1244
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1245..1347
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1348..2163
                     /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(1421..1440)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2184..2211
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2223..2810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2303..2322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2832..2861)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 acgtctcgat ggagagaccg 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 gggtctctga
     1141 ctcgagacga cgaattctct agatatcgct caatactgac catttaaatc atacctgacc
     1201 tccatagcag aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg gcacgtaaga
     1261 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     1321 agattttcag gagctaagga agctaaaatg agccatattc aacgggaaac gtcttgctcg
     1381 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1441 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     1501 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcagg
     1561 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1621 gatgatgcat ggttactcac cactgcgatc ccagggaaaa cagcattcca ggtattagaa
     1681 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     1741 cattcgattc ctgtttgtaa ttgtcctttt aacggcgatc gcgtatttcg tctcgctcag
     1801 gcgcaatcac gaatgaataa cggtttggtt ggtgcgagtg attttgatga cgagcgtaat
     1861 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     1921 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     1981 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2041 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2101 ggtattgata atcctgatat gaataaattg cagtttcact tgatgctcga tgagtttttc
     2161 tgagggccca aatgtaatca cctggctcac cttcgggtgg gcctttctgc gttgctggcg
     2221 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgatgctc aagtcagagg
     2281 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2341 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2401 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2461 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2521 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2581 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2641 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2701 acctcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2821 tgattttcta ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg attgcagtcc
     2881 agttacgctg gagtctgagg ctcgtcctga atgatatcaa gcttgaattc gtt
//
