LOCUS       sequence-249804-        2867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes pCMV-Blasticidin resistance cassette as a type 6 part to be 
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2867)
  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 2867)
  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_003
FEATURES             Location/Qualifiers
     source          1..2867
                     /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"
     promoter        1132..1335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1286..1306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1357..1366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1363..1761
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(1773..1790)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1779..1986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      complement(2019..2053)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2124..2783)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2698..2717
                     /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 gcggtgggct ctatggggta cctacgtagg tacataactt
      841 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      901 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      961 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     1021 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1081 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1141 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1201 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1261 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1321 tatataagca gagctcgttt agtgaaccgt cagatagcca ccatggccaa gcctttgtct
     1381 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     1441 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     1501 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     1561 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     1621 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     1681 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     1741 ggttatgtgt gggagggcta actcgagcga cgcctcgact gtgccttcta gttgccagcc
     1801 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1861 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1921 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1981 tggggatgcg gtgggctcta tgagttgaga ccagaccaat aaaaaacgcc cggcggcaac
     2041 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     2101 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     2161 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     2221 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     2281 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     2341 ttggctgaaa cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     2401 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     2461 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     2521 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     2581 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     2641 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     2701 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     2761 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     2821 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttgga
//
