LOCUS       sequence-271924-        3145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the F1 Kanamycin resistance cassette as a type 8a part to be
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3145)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 3145)
  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"; doi: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK8a_005
FEATURES             Location/Qualifiers
     source          1..3145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     380..399
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     464..483
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      697..1285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1186..1205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1406..1424
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1431..1449
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1467..1478
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1485..2162
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2180..2251
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2267..2294
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      2324..2752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2411..2430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2621..2642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2802..3145,1..472))
                     /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(join(2802..3145,1..472))
                     /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"
ORIGIN
        1 tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat
       61 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
      121 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
      181 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
      241 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
      301 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
      361 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
      421 tggaatttaa tcgcggcctg gagcaagacg tttcccgttg aatatggctc ataacacccc
      481 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
      541 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt
      601 ttgctgagtt gaaggatcag tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
      661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
      721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
      781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      841 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1321 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1381 ccgtgcggcc gccaattgag accagtccct atcagtgata gagattgaca tccctatcag
     1441 tgatagagat actgagcacg gatctgaaag aggagaaagg atctatggcg agtagcgaag
     1501 acgttatcaa agagttcatg cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg
     1561 agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga aggtactcag accgctaaac
     1621 tgaaagttac caaaggtggt ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc
     1681 agtacggttc caaagcttac gttaaacacc cggctgacat cccggactac ctgaaactgt
     1741 ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta
     1801 ccgttaccca ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg
     1861 gtactaactt cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt
     1921 ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga
     1981 aactgaaaga cggtggtcac tacgacgctg aagttaaaac cacctacatg gctaaaaaac
     2041 cggttcagct gccgggtgct tacaaaaccg acatcaaact ggacatcacc tcccacaacg
     2101 aagactacac catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc accggtgctt
     2161 aataaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2221 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     2281 tgggcctttc tgcgtttata agtcggtctc accgagcggc cgcacgcgcc ctgtagcggc
     2341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2401 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2521 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2641 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2701 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2761 atattaacgc ttacgtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
     2821 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2881 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2941 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     3001 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     3061 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3121 aaatcactcg catcaaccaa accgt
//
