LOCUS       sequence-249773-        2449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a nuclear localized mAzamiGreen coexpression cassette 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 2449)
  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 2449)
  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_014
FEATURES             Location/Qualifiers
     source          1..2449
                     /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..824
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             825..845
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             861..1535
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     CDS             861..1535
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     terminator      complement(1559..1593)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1664..2323)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2238..2257
                     /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 ctcaatccgg aagcggagct actaacttca
      781 gcctgctgaa gcaggctggt gacgtggagg agaaccctgg acctcccaag aagaagagga
      841 aggtggagga cgccggatcc gtgagcgtga tcaagcccga gatgaagatc aagctgtgca
      901 tgaggggcac cgtgaacggc cacaacttcg tgatcgaggg cgagggcaag ggcaacccct
      961 acgagggcac ccagatcctg gacctgaacg tgaccgaggg cgcccccctg cccttcgcct
     1021 acgacatcct gaccaccgtg ttccagtacg gcaacagggc cttcaccaag taccccgccg
     1081 acatccagga ctacttcaag cagaccttcc ccgagggcta ccactgggag aggagcatga
     1141 cctacgagga ccagggcatc tgcaccgcca ccagcaacat cagcatgagg ggcgactgct
     1201 tcttctacga catcaggttc gacggcacca acttcccccc caacggcccc gtgatgcaga
     1261 agaagaccct gaagtgggag cccagcaccg agaagatgta cgtggaggac ggcgtgctga
     1321 agggcgacgt gaacatgagg ctgctgctgg agggcggcgg ccactacagg tgcgacttca
     1381 agaccaccta caaggccaag aaggaggtga ggctgcccga cgcccacaag atcgaccaca
     1441 ggatcgagat cctgaagcac gacaaggact acaacaaggt gaagctgtac gagaacgccg
     1501 tggccaggta ctccatgctg cccagccagg cctaactcga gtggctgaga ccagaccaat
     1561 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     1621 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     1681 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     1741 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     1801 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     1861 tggtgaaact cacccaggga ttggctgaaa cgaaaaacat attctcaata aaccctttag
     1921 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     1981 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     2041 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     2101 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     2161 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     2221 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     2281 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     2341 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     2401 agttggaacc tcttacgtgc ccgatcaatc atgaccaaaa tcccttaac
//
