LOCUS       sequence-192743-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit connector-entry vector.
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: YCe2764 HC_Kan_RFP-Connector entry vector
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    84..105
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    206..227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(242..247,248..265,266..272)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    280..296
                     /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             314..325
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             332..1009
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1054..1125
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1141..1168
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1175..1195
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1278..1309
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1310..1412
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1413..2228
                     /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(1486..1505)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2249..2276
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2288..2875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2368..2387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2897..2926)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tgattgcagt ccagttacgc tggagtctga ggctcgtcct gaatgatatc aagcttgaat
       61 tcgtttttaa cgcagcagag accgaattcg cggccgcttc tagagcaata cgcaaaccgc
      121 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      181 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      241 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      301 acacatacta gagaaagagg agaaatacta gatggcttcc tccgaagacg ttatcaaaga
      361 gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga
      421 aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa
      481 aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa
      541 agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg
      601 tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga
      661 ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc
      721 gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat
      781 gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg
      841 tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc
      901 gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat
      961 cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat aacgctgata
     1021 gtgctagtgt agatcgctac tagagccagg catcaaataa aacgaaaggc tcagtcgaaa
     1081 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1141 ggctcacctt cgggtgggcc tttctgcgtt tatatactag tagcggccgc tgcagggtct
     1201 ctaggccgtt tatgacgaat tctctagata tcgctcaata ctgaccattt aaatcatacc
     1261 tgacctccat agcagaaagt caaaagcctc cgaccggagg cttttgactt gatcggcacg
     1321 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     1381 tatcgagatt ttcaggagct aaggaagcta aaatgagcca tattcaacgg gaaacgtctt
     1441 gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1501 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     1561 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1621 tcaggctaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1681 ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca ttccaggtat
     1741 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1801 ggttgcattc gattcctgtt tgtaattgtc cttttaacgg cgatcgcgta tttcgtctcg
     1861 ctcaggcgca atcacgaatg aataacggtt tggttggtgc gagtgatttt gatgacgagc
     1921 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     1981 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2041 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2101 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2161 aatatggtat tgataatcct gatatgaata aattgcagtt tcacttgatg ctcgatgagt
     2221 ttttctgagg gcccaaatgt aatcacctgg ctcaccttcg ggtgggcctt tctgcgttgc
     2281 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga tgctcaagtc
     2341 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2401 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2461 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2521 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2581 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2641 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2701 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2761 cagttacctc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2821 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2881 tcctttgatt ttctaccgaa gaaaggccca cccgtgaagg tgagccagtg agt
//
