LOCUS       sequence-306290-        6125 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Amber suppression for incorporating 3-aminotyrosine to proteins in 
            E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6125)
  AUTHORS   Zhang S, Ai HW
  TITLE     A general strategy to red-shift green fluorescent protein-based 
            biosensors.
  JOURNAL   Nat Chem Biol. 2020 Sep 14. pii: 10.1038/s41589-020-0641-7. doi: 
            10.1038/s41589-020-0641-7.
  PUBMED    32929278
REFERENCE   2  (bases 1 to 6125)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2020 Sep 14. pii: 10.1038/s41589-020-0641-7. doi: 
            10.1038/s41589-020-0641-7."
COMMENT     Sequence Label: pEvol-MjaYRS
FEATURES             Location/Qualifiers
     source          1..6125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(80..97)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2000..2659)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2574..2593
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2660..2762)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3154..3171)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3288..3833)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3368..3387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3979..4857)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4884..5168
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     5091..5110
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc tgttttggcg
       61 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
      121 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
      181 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
      241 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggccttgtt tgtgagctcc
      301 cggtcatcaa tcatccccat aatccttgtt agattatcaa ttttaaaaaa ctaacagttg
      361 tcagcctgtc ccgctttaat atcatacgcc gttatacgtt gtttacgctt tgaggaatcc
      421 catatggacg aatttgaaat gataaagaga aacacatctg aaattatcag cgaggaagag
      481 ttaagagagg ttttaaaaaa agatgaaaaa tctgcttaca taggttttga accaagtggt
      541 aaaatacatt tagggcatta tctccaaata aaaaagatga ttgatttaca aaatgctgga
      601 tttgatataa ttatattgtt ggctgattta agcgcctatt taaaccagaa aggagagttg
      661 gatgagatta gaaaaatagg agattataac aaaaaagttt ttgaagcaat ggggttaaag
      721 gcaaaatatg tttatggaag tgaattccag cttgataagg attatacact gaatgtctat
      781 agattggctt taaaaactac cttaaaaaga gcaagaagga gtatggaact tatagcaaga
      841 gaggatgaaa atccaaaggt tgctgaagtt atctatccaa taatgcaggt taatactagc
      901 cattatttag gcgttgatgt tgcagttgga gggatggagc agagaaaaat acacatgtta
      961 gcaagggagc ttttaccaaa aaaggttgtt tgtattcaca accctgtctt aacgggtttg
     1021 gatggagaag gaaagatgag ttcttcaaaa gggaatttta tagctgttga tgactctcca
     1081 gaagagatta gggctaagat aaagaaagca tactgcccag ctggagttgt tgaaggaaat
     1141 ccaataatgg agatagctaa atatttcctt gaatatcctt taaccataaa aaggccagaa
     1201 aaatttggtg gagatttgac agttaatagc tatgaggagt tagagagttt atttaaaaat
     1261 aaggaattgc atccaatgta tttaaaaaat gctgtagctg aagaacttat aaagatttta
     1321 gagccaatta gaaagagact gtaactgcag tttcaaacgc taaattgcct gatgcgctac
     1381 gcttatcagg cctacatgat ctctgcaata tattgagttt gcgtgctttt gtaggccgga
     1441 taaggcgttc acgccgcatc cggcaagaaa cagcaaacaa tccaaaacgc cgcgttcagc
     1501 ggcgtttttt ctgcttttct tcgcgaatta attccgcttc gcaacatgtg agcaccggtt
     1561 tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc
     1621 aggctctccc cgtggaggta ataattgacg atatgatcag tgcacggcta actaagcggc
     1681 ctgctgactt tctcgccgat caaaaggcat tttgctatta agggattgac gagggcgtat
     1741 ctgcgcagta agatgcgccc cgcattccgg cggtagttca gcagggcaga acggcggact
     1801 ctaaatccgc atggcagggg ttcaaatccc ctccgccgga ccaaattcga aaagcctgct
     1861 caacgagcag gcttttttgc atgctcgagc agctcagggt cgaatttgct ttcgaatttc
     1921 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     1981 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     2041 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     2101 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     2161 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     2221 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     2281 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     2341 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     2401 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     2461 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     2521 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2581 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2641 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2701 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     2761 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     2821 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt
     2881 cgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg
     2941 gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc
     3001 aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg
     3061 atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca
     3121 gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     3181 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     3241 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     3301 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     3361 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     3421 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     3481 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     3541 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     3601 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     3661 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     3721 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     3781 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     3841 atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag
     3901 cacctgaagt cagccccata cgatataagt tgtaattctc atgtttgaca gcttatcatc
     3961 gataagcttg gtacccaatt atgacaactt gacggctaca tcattcactt tttcttcaca
     4021 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
     4081 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
     4141 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
     4201 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
     4261 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
     4321 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
     4381 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
     4441 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
     4501 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
     4561 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
     4621 aatctctcct ggcgggaaca gcaaaatatc actcggtcgg caaacaaatt ctcgtccctg
     4681 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
     4741 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
     4801 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
     4861 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
     4921 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
     4981 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
     5041 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
     5101 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
     5161 ttctccatac ccgttttttt gggctaacag gaggaattag atctatggac gaatttgaaa
     5221 tgataaagag aaacacatct gaaattatca gcgaggaaga gttaagagag gttttaaaaa
     5281 aagatgaaaa atctgcttac ataggttttg aaccaagtgg taaaatacat ttagggcatt
     5341 atctccaaat aaaaaagatg attgatttac aaaatgctgg atttgatata attatattgt
     5401 tggctgattt aagcgcctat ttaaaccaga aaggagagtt ggatgagatt agaaaaatag
     5461 gagattataa caaaaaagtt tttgaagcaa tggggttaaa ggcaaaatat gtttatggaa
     5521 gtgaattcca gcttgataag gattatacac tgaatgtcta tagattggct ttaaaaacta
     5581 ccttaaaaag agcaagaagg agtatggaac ttatagcaag agaggatgaa aatccaaagg
     5641 ttgctgaagt tatctatcca ataatgcagg ttaatactag ccattattta ggcgttgatg
     5701 ttgcagttgg agggatggag cagagaaaaa tacacatgtt agcaagggag cttttaccaa
     5761 aaaaggttgt ttgtattcac aaccctgtct taacgggttt ggatggagaa ggaaagatga
     5821 gttcttcaaa agggaatttt atagctgttg atgactctcc agaagagatt agggctaaga
     5881 taaagaaagc atactgccca gctggagttg ttgaaggaaa tccaataatg gagatagcta
     5941 aatatttcct tgaatatcct ttaaccataa aaaggccaga aaaatttggt ggagatttga
     6001 cagttaatag ctatgaggag ttagagagtt tatttaaaaa taaggaattg catccaatgt
     6061 atttaaaaaa tgctgtagct gaagaactta taaagatttt agagccaatt agaaagagat
     6121 tataa
//
