LOCUS       sequence-248680-        3924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes bicistronic ABI gene expression system 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 3924)
  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 3924)
  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_027
FEATURES             Location/Qualifiers
     source          1..3924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      131..719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     620..639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             837..857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             858..1091
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1632..1658
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1671..1724
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1725..2162
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3066..3089
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(3109..3143)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3214..3873)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3788..3807
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaatcatga
       61 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      781 tctttcctgc gttatcccct gattctgtgg ataaccgtag tcggtctcat atgggcccta
      841 aaaagaagcg taaagtcgcc cccccgaccg atgtcagcct gggggacgag ctccacttag
      901 acggcgagga cgtggcgatg gcgcatgccg acgcgctaga cgatttcgat ctggacatgt
      961 tgggggacgg ggattccccg ggtccgggat ttacccccca cgactccgcc ccctacggcg
     1021 ctctggatat ggccgacttc gagtttgagc agatgtttac cgatgccctt ggaattgacg
     1081 agtacggtgg gggcgcgcca actcaagacg aattcaccca actctcccaa tcaatcgccg
     1141 agttccacac gtaccaactc ggtaacggcc gttgctcatc tctcctagct cagcgaatcc
     1201 acgcgccgcc ggaaacagta tggtccgtgg tcagacgttt cgataggcca cagatttaca
     1261 aacacttcat caaaagctgt aacgtgagtg aagatttcga gatgcgagtg ggatgcacgc
     1321 gcgacgtgaa cgtgataagt ggattaccgg cgaatacgtc acgagagaga ttagatctgt
     1381 tggacgatga tcggagagtg actgggttta gtataaccgg tggtgaacat aggctgagga
     1441 attataaatc ggttacgacg gttcatagat ttgagaaaga agaagaagaa gaaaggatct
     1501 ggaccgttgt tttggaatct tatgttgttg atgtaccgga aggtaattcg gaggaagata
     1561 cgagattgtt tgctgatacg gttattagat tgaatcttca gaaacttgct tcgatcactg
     1621 aagctatgaa ctacccatac gatgttccag attacgctag cggaagcgga gagggcagag
     1681 gaagtctgct aacatgcggt gacgtcgagg agaatcctgg acctaagcta ctgtcttcta
     1741 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
     1801 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
     1861 caccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
     1921 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1981 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     2041 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     2101 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     2161 cgacgcgtat ggtgcctttg tatggtttta cttcgatttg tggaagaaga cctgagatgg
     2221 aagctgctgt ttcgactata ccaagattcc ttcaatcttc ctctggttcg atgttagatg
     2281 gtcggtttga tcctcaatcc gccgctcatt tcttcggtgt ttacgacggc catggcggtt
     2341 ctcaggtagc gaactattgt agagagagga tgcatttggc tttggcggag gagatagcta
     2401 aggagaaacc gatgctctgc gatggtgata cgtggctgga gaagtggaag aaagctcttt
     2461 tcaactcgtt cctgagagtt gactcggaga ttgagtcagt tgcgccggag acagttgggt
     2521 caacgtcggt ggttgccgtt gttttcccgt cacacatctt cgtcgctaac tgcggtgact
     2581 ctagagccgt tctttgccgc ggcaaaactg cacttccatt atccgttgac cataaaccgg
     2641 atagagaaga tgaagctgcg aggattgaag ccgcaggagg gaaagtgatt cagtggaatg
     2701 gagctcgtgt tttcggtgtt ctcgccatgt cgagatccat tggcgataga tacttgaaac
     2761 catccatcat tcctgatccg gaagtgacgg ctgtgaagag agtaaaagaa gatgattgtc
     2821 tgattttggc gagtgacggg gtttgggatg taatgacgga tgaagaagcg tgtgagatgg
     2881 caaggaagcg gattctcttg tggcacaaga aaaacgcggt ggctggggat gcatcgttgc
     2941 tcgcggatga gcggagaaag gaagggaaag atcctgcggc gatgtccgcg gctgagtatt
     3001 tgtcaaagct ggcgatacag agaggaagca aagacaacat aagtgtggtg gtggttgatt
     3061 tgaaggatta caaggacgat gacgataagg gatcctgaga ccagaccaat aaaaaacgcc
     3121 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
     3181 atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca
     3241 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     3301 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     3361 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     3421 cacccaggga ttggctgaaa cgaaaaacat attctcaata aaccctttag ggaaataggc
     3481 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     3541 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     3601 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc
     3661 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     3721 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     3781 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     3841 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     3901 cgataactca aaaaatacgc ccgg
//
