LOCUS       sequence-249068-        2682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Gibberellin Insensitive tagged with his and tagBFP as a Type
            3a part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2682)
  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 2682)
  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: MTK3a_022
FEATURES             Location/Qualifiers
     source          1..2682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      56..644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     545..564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             768..785
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1059..1757
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     terminator      complement(1792..1826)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1897..2556)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2471..2490
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
       61 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      721 cccctgattc tgtggataac cgtagtcggt ctcatatgaa gagagatcat catcatcatc
      781 atcatcaaga taagaagact atgatgatga atgaagaaga cgacggtaac ggcatggatg
      841 agcttctagc tgttcttggt tacaaggtta ggtcatccga aatggctgat gttgctcaga
      901 aactcgagca gcttgaagtt atgatgtcta atgttcaaga agacgatctt tctcaactcg
      961 ctactgagac tgttcactat aatccggcgg agctttacac gtggcttgat tctatgctca
     1021 ccgaccttaa tcatatgtca agcgctgcag gtgctaccat gagcgagctg attaaggaga
     1081 acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat
     1141 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
     1201 gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga
     1261 ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct
     1321 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     1381 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat
     1441 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc gaaacgctgt
     1501 accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga
     1561 gccatctgat cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca
     1621 agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg
     1681 aaacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac
     1741 tggggcacaa gcttaattca agcgctgcag gtggttcttg agaccagacc aataaaaaac
     1801 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     1861 tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc
     1921 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     1981 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     2041 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     2101 actcacccag ggattggctg aaacgaaaaa catattctca ataaaccctt tagggaaata
     2161 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     2221 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     2281 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa
     2341 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     2401 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     2461 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     2521 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     2581 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     2641 acctcttacg tgcccgatca atcatgacca aaatccctta ac
//
