LOCUS       sequence-249973-        3115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes promoter-less tagBFP T2A Blasticidin resistance cassette to 
            be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3115)
  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 3115)
  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: MTK7_010
FEATURES             Location/Qualifiers
     source          1..3115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             714..1412
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1413..1433
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1464..1517
                     /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             1524..1922
                     /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(1934..1951)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1940..2147
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      complement(2180..2214)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2285..2944)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2859..2878
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 tctttcctgc gttatcccct gattctgtgg ataaccgtag tcggtctcag agtatgagcg
      721 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc
      781 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
      841 tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc
      901 tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt
      961 ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga
     1021 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
     1081 gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
     1141 tcaccgaaac gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga
     1201 agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac
     1261 ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca
     1321 aggaggccaa caacgaaacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg
     1381 acctccctag caaactgggg cacaagctta atcccaagaa gaagaggaag gtggaggacg
     1441 cccgtacgag tactggaagc ggagagggca gaggaagtct gctaacatgc ggtgacgtcg
     1501 aggagaatcc tggacctgga tccatggcca agcctttgtc tcaagaagaa tccaccctca
     1561 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     1621 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     1681 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     1741 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     1801 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     1861 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     1921 aactcgagcg acgcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1981 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2041 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2101 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2161 atccgatgag accagaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     2221 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat
     2281 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     2341 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     2401 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     2461 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgaa acgaaaaaca
     2521 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     2581 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     2641 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2701 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     2761 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     2821 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     2881 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     2941 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     3001 atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaat catgaccaaa
     3061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatca
//
