LOCUS       sequence-249791-        2059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ha-tagged and nuclear localized Gibberellin Insensitive as a
            Type 4a part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2059)
  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 2059)
  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: MTK4a_027
FEATURES             Location/Qualifiers
     source          1..2059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      113..701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     602..621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             819..845
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             825..851
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             864..884
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             891..911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      complement(1226..1260)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1331..1990)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1905..1924
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 atggtgaaag ttggaacctc ttacgtgccc gatcaatcat gaccaaaatc ccttaacgtg
       61 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
      781 ctgattctgt ggataaccgt agtcggtctc aatccgccta tccctatgac gtgcccgatt
      841 atgccagcct gggcagcggc tcccccaaga aaaaacgcaa ggtggaagat cctaagaaaa
      901 agcggaaagt ggacggcatt ggtagtggga gcaacggagg tggaggctct ggacctaggc
      961 aagataagaa gactatgatg atgaatgaag aagacgacgg taacggcatg gatgagcttc
     1021 tagctgttct tggttacaag gttaggtcat ccgaaatggc tgatgttgct cagaaactcg
     1081 agcagcttga agttatgatg tctaatgttc aagaagacga tctttctcaa ctcgctactg
     1141 agactgttca ctataatccg gcggagcttt acacgtggct tgattctatg ctcaccgacc
     1201 ttaattaatg gctgagacca gaccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     1261 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     1321 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     1381 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     1441 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     1501 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga
     1561 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     1621 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     1681 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     1741 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
     1801 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     1861 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     1921 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     1981 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     2041 gtagtgatct tatttcatt
//
