LOCUS       sequence-248659-        2661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes HA-tagged PIF6 fused to Tet Repressor protein as a Type 3 
            part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2661)
  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 2661)
  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: MTK3_011
FEATURES             Location/Qualifiers
     source          1..2661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             777..1397
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(850..869)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1746..1772
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      complement(1792..1826)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1897..2556)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2471..2490
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg tctcatatgt
      781 ctagattaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa
      841 tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt
      901 attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata
      961 ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata
     1021 acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag
     1081 gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc
     1141 aacaaggttt ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt
     1201 taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta
     1261 ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg
     1321 cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta
     1381 aatgtgaaag tgggtccgcg tacagccgcg cgctcgatag tgctggtagt gctggtagtg
     1441 ctggtatgat gttcttacca accgattatt gttgcaggtt aagcgatcaa gagtatatgg
     1501 agcttgtgtt tgagaatggc cagattcttg caaagggcca aagatccaac gtttctctgc
     1561 ataatcaacg taccaaatcg atcatggatt tgtatgaggc agagtataac gaggatttca
     1621 tgaagagtat catccatggt ggtggtggtg ccatcacaaa tctcggggac acgcaggttg
     1681 ttccacaaag tcatgttgct gctgcccatg aaacaaacat gttggaaagc aataaacatg
     1741 ttgactaccc atacgatgtt ccagattacg ctggatcctg agaccagacc aataaaaaac
     1801 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     1861 tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc
     1921 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     1981 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     2041 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     2101 actcacccag ggattggctg aaacgaaaaa catattctca ataaaccctt tagggaaata
     2161 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     2221 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     2281 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa
     2341 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     2401 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     2461 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     2521 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     2581 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     2641 acctcttacg tgcccgatca a
//
