LOCUS       sequence-110250-        2694 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes KanR-ColE1 as a Type 8a part to be used in the Dueber YTK 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2694)
  AUTHORS   Lee ME, DeLoache WC, Cervantes B, Dueber JE
  TITLE     A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
  JOURNAL   ACS Synth Biol. 2015 Apr 14.
  PUBMED    25871405
REFERENCE   2  (bases 1 to 2694)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2015 Apr 14."
COMMENT     Sequence Label: pYTK090
FEATURES             Location/Qualifiers
     source          1..2694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..864)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(49..864)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     772..791
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     856..875
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1089..1677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1578..1597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1798..1816
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1823..1841
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1859..1870
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1877..2554
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2572..2643
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2659..2686
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tcggtctcac cgagcggccg cgtgttacaa ccaattaacc aattctgatt agaaaaactc
       61 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      121 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
      181 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
      241 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
      301 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
      361 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
      421 gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
      481 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
      541 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
      601 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
      661 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
      721 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
      781 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga
      841 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
      901 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
      961 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agtcatgacc
     1021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1201 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1741 tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg ccgccaattg agaccagtcc
     1801 ctatcagtga tagagattga catccctatc agtgatagag atactgagca cggatctgaa
     1861 agaggagaaa ggatctatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa
     1921 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg
     1981 tcgtccgtac gaaggtactc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc
     2041 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca
     2101 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg
     2161 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga
     2221 cggtgagttc atctacaaag ttaaactgcg tggtactaac ttcccgtccg acggtccggt
     2281 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg
     2341 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc
     2401 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac
     2461 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga
     2521 acgtgctgaa ggtcgtcact ccaccggtgc ttaataagga tctccaggca tcaaataaaa
     2581 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2641 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taag
//
