LOCUS       sequence-249737-        2865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Kruppel Associated Box repression domain fused to methyl CpG
            binding protein 2 as a Type 3b part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2865)
  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 2865)
  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: MTK3b_031
FEATURES             Location/Qualifiers
     source          1..2865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     408..427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             622..642
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             703..897
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             931..951
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             961..1818
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     terminator      complement(1838..1872)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1943..2602)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2517..2536
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2784..2865,1..507)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
       61 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      601 aaccgtagtc ggtctcattc tcccaagaag aagaggaagg tgagtggtgg aggaagtggc
      661 gggtcagggt cgatggacgc gaaatcactt acggcatggt cgagaacact ggttacgttc
      721 aaggacgtgt ttgtggactt tacacgtgag gagtggaaat tgctggatac tgcgcaacaa
      781 attgtgtatc gaaatgtcat gcttgagaat tacaagaacc tcgtcagtct cggataccag
      841 ttgacgaaac cggatgtgat ccttaggctc gaaaaggggg aagaaccttg gctggtatcg
      901 ggaggtggtt cgggtggctc tggatcaagc ccaaagaaga aacggaaggt ggaagcctca
      961 gtgcaggtga aaagggtgct ggaaaaatcc cccggcaaac tcctcgtgaa gatgcccttc
     1021 caggcttccc ctggcggaaa aggtgaaggg ggtggcgcaa ccacatctgc ccaggtcatg
     1081 gtcatcaagc gacctggaag gaaaagaaag gccgaggctg accctcaggc cattccaaag
     1141 aaacggggac gcaagccagg gtccgtggtc gcagctgcag cagctgaggc taagaaaaag
     1201 gcagtgaagg aaagctccat ccgcagtgtg caggagactg tcctgcccat caagaagagg
     1261 aagactaggg aaaccgtgtc catcgaggtc aaagaagtgg tcaagcccct gctcgtgtcc
     1321 accctgggcg aaaaatctgg aaaggggctc aaaacatgca agtcacctgg acggaaaagc
     1381 aaggagtcta gtccaaaggg gcgctcaagc tccgcttcta gtccccctaa aaaggaacac
     1441 catcaccatc accatcacgc cgagtctcct aaggctccta tgccactgct cccaccacct
     1501 ccaccacctg agccacagtc aagcgaagac cccatcagcc cacccgagcc tcaggatctg
     1561 tcctctagta tttgcaaaga ggaaaagatg cccagagcag gcagcctgga gagtgatggc
     1621 tgtccaaaag aacccgccaa gacccagcct atggtggcag ccgctgcaac taccaccaca
     1681 accacaacta ccacagtggc cgaaaaatac aagcatcgcg gcgagggcga acgaaaggac
     1741 attgtgtcaa gctccatgcc cagacctaac cgggaggaac cagtcgatag taggacaccc
     1801 gtgactgaga gagtctcagg atcctgagac cagaccaata aaaaacgccc ggcggcaacc
     1861 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     1921 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     1981 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2041 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2101 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2161 tggctgaaac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2221 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2281 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2341 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     2401 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     2461 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     2521 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     2581 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     2641 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     2701 cgatcaatca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggat
//
