LOCUS       sequence-271981-        5299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the hCLYBL landing pad destination vector as a type 0 part 
            to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5299)
  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 5299)
  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: MTK0_050
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(670..1485)
                     /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(670..1485)
                     /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     1393..1412
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1477..1496
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1710..2298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2199..2218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3368..3401
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     RBS             3708..3719
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3726..4439
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4451..4522
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4538..4565
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    4938..4971
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ttcctgtctc agcctaccaa gtagctgaga ctataggtgc aaaaaagcca ctatacctgg
       61 ctagtttaca ggttttaaca aatgcattat gccacgtatc cattattaca ggatcacaca
      121 agatattttc attaccctga aagcatccct gtgttccacc aattcatcct gcctccatga
      181 gccgctggca accactgatc tctatagttt tgccttttct aaaatgtcat ataattggaa
      241 tcatacagtc tgtagcattt tcagactagc ttttaaaatt tggcaatatg catttaaggt
      301 tcctccttaa atgtgaaggg catagccaat gtggcttgat agctcatttc tttttattgg
      361 tgaatatttc attgtctgga tgttccacag tttgtttatc cattcaccta ttcaatttgc
      421 tttttttctg tgtatctatc actaattcaa tactggactc tccaacagag cccacttata
      481 tgaggtacct ggagtagtca gactgataga gacagaaact agattggtgg ttgccagggg
      541 cagagaggaa ggggaatgag gagttattgc ttcaagaata caaagtttca ctttttaaga
      601 tgaaaggttt tctggaggta gatggtggtg ccgagcggcc gcgtgttaca accaattaac
      661 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
      721 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
      781 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
      841 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
      901 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
      961 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     1021 tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac
     1081 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     1141 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     1201 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     1261 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     1321 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     1381 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     1441 taatcgcggc ctggagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     1501 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     1561 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     1621 gttgaaggat cagtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1681 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1741 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1801 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1861 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1921 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1981 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2041 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2101 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2161 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2221 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2281 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2341 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtgcg
     2401 gccgccaata gccttatacc tcctcatctc aaaacttatt acaaaaccac agttttcaaa
     2461 acagtgtggt actgcacata tatagaccaa tggactagga tagagagccc ataagtaaac
     2521 ccttgtatgt atggtcagca ggagaatcat gccaatgggc caatatacat tctgacccac
     2581 agtttcataa taaaataaaa tggttgtggt tgtaagccac tatgtttcag agtggtttgt
     2641 tacacagcaa taaataacta atatagtagg cataccatca agtccaaagt aggtagagaa
     2701 gaatgtaaat agcagagcaa aacagcatga ctggtggctg ggaggcttaa aactgggaca
     2761 ggatcagagt catgaaagaa gtcaaagaaa tggttcagaa gtaaggctga gactgactta
     2821 caaaagctga aagtcccttt aagttggtgt ttggtgcatt ggcaggggca ggtatggtga
     2881 cttaaaagag ccatgctcaa caagatcaag cacaacacaa tcacgggtca ccccagcaga
     2941 ccttagcgag tctagccatt tctttggtgg tggtcacagt catgcttcag cccagtttcc
     3001 acttggacaa atggtacata ttttcaatga gatgaaaatt aagatacaat ccatgtgctc
     3061 agagagtgat cacagctctg actaaacact gtgccccaaa gtgttgagga attgggaaaa
     3121 cctagctgag ttagtggtct gttttctgtt acaataaagc tcataatgaa aattagcctt
     3181 ctttgttctt ccccaagtct ttctttctag acgaaactac tttcaactgt tttaacttcc
     3241 ttactgttaa cttccatatt tctagatagt atggtcagac tgttatttct tgacttttaa
     3301 atgtaagata ttatctactg acttccttct atgtaagatg aggattgagc tctcttaccc
     3361 ttcccctgaa gttcctattc tctagaaagt ataggaactt cctgatgaga cgtggtagag
     3421 ccacaaacag ccggtacaag caacgatctc caggaccatc tgaatcatgc gcggatgaca
     3481 cgaactcacg acggcgatca cagacattaa cccacagtac agacactgcg acaacgtggc
     3541 aattcgtcgc aatacaacgg aaagtgaaac gtgatttcat gcgtcatttt gaacattttg
     3601 taaatcttat ttaataatgt gtgcggcaat tcacatttaa tttatgaatg ttttcttaac
     3661 atcgcggcaa ctcaagaaac ggcaggttcg gatcttagct actagagaaa gaggagaaat
     3721 actagatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
     3781 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
     3841 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggtt ccttggccga
     3901 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
     3961 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
     4021 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
     4081 tggtaaaccg cattgagctg aaaggcattg actttaaaga ggacggcaat atcctgggcc
     4141 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
     4201 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
     4261 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
     4321 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
     4381 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat
     4441 gaccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     4501 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     4561 ctgcgtttat agctggaaat ctgctcgtca gtggtgctca cactgacgaa tcatgtacag
     4621 atcataccga tgactgcctg gcgactcaca actaagcaag acagccggaa ccagcgccgg
     4681 cgaacaccac tgcatatatg gcatatcaca acagtccacg tctcaagcag ttacagagat
     4741 gttacgaacc ctgaacaggt tgactattaa ttgtgtctgc ttgatgtgga caccaggtgg
     4801 cgctggacat cagatttgga gaggcagttg tctagggaac cgggctctgt gccagcgcag
     4861 gaggcaggct ggctctccta ttccagggat gctcatccag gaaggaaagg ttgcatgctg
     4921 gacacactaa ccttacagaa gttcctattc tctagaaagt ataggaactt cgagttcctc
     4981 atccttccaa cataaatata ttttgggatt atatcaacat tcaatgttac ttaaagtgac
     5041 cttgtaaata ttttcacaac tgagccatgt ttgatttgta tacttatgtt tactttactg
     5101 tttttcctga agttaataat tgccttgaat ttatttattt ctttaaaaat gtttcattac
     5161 tcaggactgt agtttacatt acgattcttt gtgttataca gttgatgggt ttcttttctt
     5221 tcttaatttc tttaaaaaat agagatgggg tcttactata ttacccaggc tggtcttgaa
     5281 gtcctgggct caagtgatc
//
