LOCUS       sequence-249818-        3206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Woodchuck Hepatitis Virus (WHP) Posttranscriptional 
            Regulatory Element (WPRE) expressing a Blasticidin resistance 
            cassette without a terminator to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3206)
  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 3206)
  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: MTK6_013
FEATURES             Location/Qualifiers
     source          1..3206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      98..686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     587..606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(855..875)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1273..1284)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        1711..1914
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1865..1885
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1936..1945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1942..2340
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     terminator      complement(2358..2392)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2463..3122)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3037..3056
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 acctcttacg tgcccgatca atcatgacca aaatccctta acgtgagttt tcgttccact
       61 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      121 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      181 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      241 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      301 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      361 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      421 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      481 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      541 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      601 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      661 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      721 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      781 accgtagtcg gtctcataca caatcaacct ctggattaca aaatttgtga aagattgact
      841 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
      901 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
      961 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     1021 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     1081 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     1141 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     1201 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     1261 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     1321 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcg
     1381 cggtgggctc tatggggtac ctacgtaggt acataactta cggtaaatgg cccgcctggc
     1441 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1501 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1561 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1621 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1681 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     1741 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     1801 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     1861 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     1921 gtgaaccgtc agatagccac catggccaag cctttgtctc aagaagaatc caccctcatt
     1981 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     2041 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     2101 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     2161 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     2221 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     2281 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     2341 gagttgagac cagaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     2401 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca
     2461 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     2521 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     2581 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     2641 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgaaac gaaaaacata
     2701 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     2761 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     2821 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     2881 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
     2941 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     3001 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     3061 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     3121 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     3181 cttatttcat tatggtgaaa gttgga
//
