LOCUS       sequence-189541-        2939 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 2939)
  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 2939)
  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..2939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      73..102
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      124..711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     612..631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(723..750)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(774..1589)
                     /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     1497..1516
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1590..1692)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1693..1724)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     misc_feature    1808..1829
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    1930..1951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1966..1971,1972..1989,1990..1996)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2004..2020
                     /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             2038..2049
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2056..2733
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2778..2849
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2865..2892
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2899..2919
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 aacgaattca agcttgatat cattcaggac gagcctcaga ctccagcgta actggactgc
       61 aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc
      121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc
      241 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      661 cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      721 aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc tcattagaaa
      781 aactcatcga gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat
      841 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
      901 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
      961 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1021 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     1081 cgctcgtcat caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga
     1141 gcgagacgaa atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac
     1201 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1261 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     1321 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg
     1381 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     1441 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     1501 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
     1561 caagacgttt cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat
     1621 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     1681 acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc
     1741 aggtatgatt taaatggtca gtattgagcg atatctagag aattcgtcac gtctcgctac
     1801 agagaccgaa ttcgcggccg cttctagagc aatacgcaaa ccgcctctcc ccgcgcgttg
     1861 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1921 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     1981 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacat actagagaaa
     2041 gaggagaaat actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa
     2101 gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt
     2161 cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg
     2221 ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac
     2281 ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt
     2341 gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac
     2401 ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt
     2461 atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt
     2521 gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct
     2581 gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc
     2641 gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa
     2701 cgtgctgaag gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg
     2761 ctactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2821 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2881 ggcctttctg cgtttatata ctagtagcgg ccgctgcagg gtctctgcaa cgagacgaa
//
