LOCUS       sequence-249071-        3336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Absisic acid insensitive 1 taggeed 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 3336)
  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 3336)
  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_023
FEATURES             Location/Qualifiers
     source          1..3336
                     /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             1713..2411
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     terminator      complement(2446..2480)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2551..3210)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3125..3144
                     /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 atcatacgcg tgtgcctttg tatggtttta cttcgatttg tggaagaaga cctgagatgg
      841 aagctgctgt ttcgactata ccaagattcc ttcaatcttc ctctggttcg atgttagatg
      901 gtcggtttga tcctcaatcc gccgctcatt tcttcggtgt ttacgacggc catggcggtt
      961 ctcaggtagc gaactattgt agagagagga tgcatttggc tttggcggag gagatagcta
     1021 aggagaaacc gatgctctgc gatggtgata cgtggctgga gaagtggaag aaagctcttt
     1081 tcaactcgtt cctgagagtt gactcggaga ttgagtcagt tgcgccggaa acggttgggt
     1141 caacgtcggt ggttgccgtt gttttcccat ctcacatctt cgtcgctaac tgcggtgact
     1201 ctagagccgt tctttgccgc ggcaaaactg cacttccatt atccgttgac cataaaccgg
     1261 atagagaaga tgaagctgcg aggattgaag ccgcaggagg gaaagtgatt cagtggaatg
     1321 gagctcgtgt tttcggtgtt ctcgccatgt cgagatccat tggcgataga tacttgaaac
     1381 catccatcat tcctgatccg gaagtgacgg ctgtgaagag agtaaaagaa gatgattgtc
     1441 tgattttggc gagtgacggg gtttgggatg taatgacgga tgaagaagcg tgtgagatgg
     1501 caaggaagcg gattctcttg tggcacaaga aaaacgcggt ggctggggat gcatcgttgc
     1561 tcgcggatga gcggagaaag gaagggaaag atcctgcggc gatgtccgcg gctgagtatt
     1621 tgtcaaagct ggcgatacag agaggaagca aagacaacat aagtgtggtg gtggttgatt
     1681 tgaagcatat gtcaagcgct gcaggtgcta ccatgagcga gctgattaag gagaacatgc
     1741 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
     1801 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
     1861 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
     1921 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
     1981 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     2041 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     2101 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgaaacg ctgtaccccg
     2161 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     2221 tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     2281 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgaaacct
     2341 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     2401 acaagcttaa ttcaagcgct gcaggtggtt cttgagacca gaccaataaa aaacgcccgg
     2461 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     2521 aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta
     2581 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     2641 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     2701 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     2761 ccagggattg gctgaaacga aaaacatatt ctcaataaac cctttaggga aataggccag
     2821 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     2881 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     2941 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg
     3001 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     3061 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     3121 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     3181 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     3241 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     3301 tacgtgcccg atcaatcatg accaaaatcc cttaac
//
