LOCUS       sequence-250330-        5299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the hThal5.10-2_hg18 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_049
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(671..1486)
                     /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(671..1486)
                     /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     1394..1413
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1478..1497
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1711..2299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2200..2219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3369..3402
                     /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             3709..3720
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3727..4440
                     /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      4452..4523
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4539..4566
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    4939..4972
                     /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 tgctttttgc tgttgtaaca gtaattcctg caaagtacat actaaaccat gtaaaattac
       61 aatcctgact attgagttag tgaattatgt atactttgag gaaagcggaa aatgtgtatt
      121 cagatagtat gtaagatagc tgcgctttct ggaggcttat agatctacat attgcctatg
      181 taatcactcg aggaaggaaa ccaagtgctc ttgcccttct aatgaatttt gaatgcatca
      241 gtaatcactg tattatgtat ctccaattaa ctagagaaac aggctcacaa tattctaaat
      301 ttgtacacta aggaagtaca taacactgtg ttgttaattg agaaaatact cattccaaca
      361 aggttaacaa ccctaaagga gttaaaattt atcaagtgct tgaacaatgc atagcactta
      421 ttaagtgcta aacaatttgt tgacaattaa atattcactg tttgaatttt gtaaattgca
      481 ttttcttcat aaaacactgc ataaatttgt ctcatatttt aaattatgac tttccttttt
      541 ttcttcaatg agtctaaata acattggtgg tgttagatat gattgaactc tattattgat
      601 tgaattcact aagcaaatta tccagctttt tccgagcggc cgcgtgttac aaccaattaa
      661 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
      721 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
      781 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
      841 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
      901 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt
      961 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1021 ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa
     1081 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1141 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1201 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1261 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1321 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1381 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1441 ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     1501 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     1561 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
     1621 agttgaagga tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2341 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtgc
     2401 ggccgccaat tagaaatgta tgaattaaat tggattaatc acaatatata ctatataaaa
     2461 aagtatgcag tacataaata cacagataca aaatataggc atcatttttg aagttatgtt
     2521 ataatatatt gaggctattt tattagaaaa cgaactttat gctaactgta taaaggaaaa
     2581 aagaataatt gcagaagaaa taatttataa tttgcttttc cttggtttta tttaaaaatt
     2641 aaattaatta aattataagt atattgtttc catataattt ataataaata aggcatatag
     2701 gccaggcacg gtggctcatg cctgtaatcc cagcactttg ggaggcagag gtgggtgaac
     2761 gcctgaggtc aggcgttcaa gaccagcctg gccaacatgg tgaaacctcg tctgtacaaa
     2821 aaatacaaaa atcagctggg catggtgtca gatgcctgta atcccaggta cttgggaggc
     2881 tgaggcagga gaatctcttg aacccaggag gcagaggttg cagtgagccg agatcatgcc
     2941 attgcactcc tgcctggggg acaagagcga gacttcgtct aaaaaacaaa acaaaacaaa
     3001 acaaaaacaa aaaaaaacac aaagaaacaa aaaaccattt aaagcaacaa attactcatt
     3061 tcttagctat attctgttta attaaaaaat agggtatcct ctatgtaaag aaaaatataa
     3121 tgttattgag ccattttctt cagaagaaca agctactcta cctatgtaga gtgataatgt
     3181 atcctcagga ttgaagatca aaagaggtaa tatatacaaa cattctttga aaggtgcctt
     3241 ataaaacaaa aaattgtttt atcataaaaa tttaaggtag tgcagagagc taacatgaaa
     3301 aatgtaaaca ttggcaataa gaacaaagat atactatgtt tgaaaaatgg ttgaagtccc
     3361 tttcccctga agttcctatt ctctagaaag tataggaact tcctgatgag acgtggtaga
     3421 gccacaaaca gccggtacaa gcaacgatct ccaggaccat ctgaatcatg cgcggatgac
     3481 acgaactcac gacggcgatc acagacatta acccacagta cagacactgc gacaacgtgg
     3541 caattcgtcg caatacaacg gaaagtgaaa cgtgatttca tgcgtcattt tgaacatttt
     3601 gtaaatctta tttaataatg tgtgcggcaa ttcacattta atttatgaat gttttcttaa
     3661 catcgcggca actcaagaaa cggcaggttc ggatcttagc tactagagaa agaggagaaa
     3721 tactagatgc gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg
     3781 gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact
     3841 aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt tccttggccg
     3901 actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg
     3961 aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt
     4021 tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc
     4081 ctggtaaacc gcattgagct gaaaggcatt gactttaaag aggacggcaa tatcctgggc
     4141 cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa
     4201 aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg
     4261 gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat
     4321 cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg
     4381 gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa
     4441 tgaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4501 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     4561 tctgcgttta tagctggaaa tctgctcgtc agtggtgctc acactgacga atcatgtaca
     4621 gatcataccg atgactgcct ggcgactcac aactaagcaa gacagccgga accagcgccg
     4681 gcgaacacca ctgcatatat ggcatatcac aacagtccac gtctcaagca gttacagaga
     4741 tgttacgaac cctgaacagg ttgactatta attgtgtctg cttgatgtgg acaccaggtg
     4801 gcgctggaca tcagatttgg agaggcagtt gtctagggaa ccgggctctg tgccagcgca
     4861 ggaggcaggc tggctctcct attccaggga tgctcatcca ggaaggaaag gttgcatgct
     4921 ggacacacta accttacaga agttcctatt ctctagaaag tataggaact tcgagtgttt
     4981 atacagttgc cttacaaaga cagtattaga ataaaaaaag cctcaggcac ttctattata
     5041 ataatgaatg tctcatgaga tgggatgatg tgcaggttac aacattctga tagtaatttc
     5101 taatgcatta cagtagagca tggactcttg agaatcagat gaaattttct tccatctaga
     5161 ttacattttc tcactgtccc aatgtgcttg atatgaactg acaccaccat aaatactttt
     5221 atctgaatgg cagcactgtc atagttagac agagaaatat atagcgttga gtgaaaaata
     5281 attttagata ctttccaag
//
