LOCUS       sequence-249089-        2340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mAzamiGreen as a Type 3b part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2340)
  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 2340)
  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: MTK3b_004
FEATURES             Location/Qualifiers
     source          1..2340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      180..768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     669..688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             883..1554
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     CDS             883..1554
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     terminator      complement(1574..1608)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1679..2338)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2253..2272
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
       61 tttcattatg gtgaaagttg gaacctctta cgtgcccgat caatcatgac caaaatccct
      121 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      181 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      241 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      301 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
      361 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      421 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      481 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      541 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      601 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      661 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      721 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      781 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      841 ttatcccctg attctgtgga taaccgtagt cggtctcatt ctgtgagcgt gatcaagccc
      901 gagatgaaga tcaagctgtg catgaggggc accgtgaacg gccacaactt cgtgatcgag
      961 ggcgagggca agggcaaccc ctacgagggc acccagatcc tggacctgaa cgtgaccgag
     1021 ggcgcccccc tgcccttcgc ctacgacatc ctgaccaccg tgttccagta cggcaacagg
     1081 gccttcacca agtaccccgc cgacatccag gactacttca agcagacctt ccccgagggc
     1141 taccactggg agaggagcat gacctacgag gaccagggca tctgcaccgc caccagcaac
     1201 atcagcatga ggggcgactg cttcttctac gacatcaggt tcgacggcac caacttcccc
     1261 cccaacggcc ccgtgatgca gaagaagacc ctgaagtggg agcccagcac cgagaagatg
     1321 tacgtggagg acggcgtgct gaagggcgac gtgaacatga ggctgctgct ggagggcggc
     1381 ggccactaca ggtgcgactt caagaccacc tacaaggcca agaaggaggt gaggctgccc
     1441 gacgcccaca agatcgacca caggatcgag atcctgaagc acgacaagga ctacaacaag
     1501 gtgaagctgt acgagaacgc cgtggccagg tactccatgc tgcccagcca ggccggatcc
     1561 tgagaccaga ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     1621 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     1681 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     1741 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     1801 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     1861 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct
     1921 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     1981 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     2041 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     2101 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     2161 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     2221 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     2281 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
//
