LOCUS       sequence-192733-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit part-entry vector position 22.
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: YCe1514 HC_Kan_RFP-p22
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    48..68
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(75..102)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(118..189)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(234..911)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPVDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             918..929
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    947..963
                     /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(971..977,978..995,996..1001))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1016..1037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    1138..1159
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1242..1273
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1274..1376
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1377..2192
                     /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(1450..1469)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2213..2240
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2252..2839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2332..2351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2861..2890)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tcctgaatga tatcaagctt gaattcgttc gtctcgttcg agagaccctg cagcggccgc
       61 tactagtata taaacgcaga aaggcccacc cgaaggtgag ccagtgtgac tctagtagag
      121 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
      181 ttttatttga tgcctggctc tagtagcgat ctacactagc actatcagcg ttattaagca
      241 ccggtggagt gacgaccttc agcacgttcg tactgttcaa cgatggtgta gtcttcgttg
      301 tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac ccggcagctg aaccggtttt
      361 ttagccatgt aggtggtttt aacttcagcg tcgtagtgac caccgtcttt cagtttcaga
      421 cgcattttga tttcaccttt cagagcaccg tctaccgggt acatacgttc ggtggaagct
      481 tcccaaccca tggttttttt ctgcataacc ggaccgtcgg acgggaagtt ggtaccacgc
      541 agtttaactt tgtagatgaa ctcaccgtct tgcagggagg agtcctgggt aacggtaaca
      601 acaccaccgt cttcgaagtt cataacacgt tcccatttga aaccttccgg gaaggacagt
      661 ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag ctttggaacc gtactggaac
      721 tgcggggaca ggatgtccca agcgaacggc agcggaccac ctttggtaac tttcagttta
      781 gcggtctggg taccttcgta cggacgacct tcaccttcac cttcgatttc gaactcgtga
      841 ccgttaacgg aaccttccat acgaactttg aaacgcatga actctttgat aacgtcttcg
      901 gaggaagcca tctagtattt ctcctctttc tctagtatgt gtgaaattgt tatccgctca
      961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgctc
     1141 tagaagcggc cgcgaattcg gtctctcaac cgagacgtac gaattctcta gatatcgctc
     1201 aatactgacc atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg
     1261 gaggcttttg acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     1321 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga
     1381 gccatattca acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg
     1441 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     1501 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     1561 ccaatgatgt tacagatgag atggtcaggc taaactggct gacggaattt atgcctcttc
     1621 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     1681 cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1741 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1801 acggcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1861 gtgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1921 tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     1981 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2041 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2101 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2161 agtttcactt gatgctcgat gagtttttct gagggcccaa atgtaatcac ctggctcacc
     2221 ttcgggtggg cctttctgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2281 atcacaaaaa tcgatgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2341 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2401 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2461 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2521 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2581 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2641 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2701 ttggtatctg cgctctgctg aagccagtta cctcggaaaa agagttggta gctcttgatc
     2761 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2821 cagaaaaaaa ggatctcaag aagatccttt gattttctac cgaagaaagg cccacccgtg
     2881 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcg
//
