LOCUS       Exported                4977 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses Mj aaRS for sulfotyrosine and Mj tRNA for decoding UAG 
            codons.
ACCESSION   .
VERSION     .
KEYWORDS    pUltra-sY
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4977)
  AUTHORS   Schwessinger B, Li X, Ellinghaus TL, Chan LJ, Wei T, Joe A, Thomas 
            N, Pruitt R, Adams PD, Chern MS, Petzold CJ, Liu CC, Ronald PC
  TITLE     A second-generation expression system for tyrosine-sulfated proteins
            and its application in crop protection.
  JOURNAL   Integr Biol (Camb). 2016 Apr 18;8(4):542-5. doi: 10.1039/c5ib00232j.
            Epub 2015 Nov 27.
  PUBMED    26611838
REFERENCE   2  (bases 1 to 4977)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4977
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        122..150
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    158..174
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(1179..1196)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     regulatory      1820..1829
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(2012..2803)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2804..2895)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2943..3681)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3891..4973)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4935..4954
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4974..74)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
       61 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttagggagct
      121 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
      181 aaaggaggtg cggccgcatg gacgaatttg aaatgataaa gagaaacaca tctgaaatta
      241 tcagcgagga agagttaaga gaggttttaa aaaaagatga aaaatctgct ctgataggtt
      301 ttgaaccaag tggtaaaata catttagggc attatctcca aataaaaaag atgattgatt
      361 tacaaaatgc tggatttgat ataattatac ctttggctga tttacatgcc tatttaaacc
      421 agaaaggaga gttggatgag attagaaaaa taggagatta taacaaaaaa gtttttgaag
      481 caatggggtt aaaggcaaaa tatgtttatg gaagtgaatt ccagcttgat aaggattata
      541 cactgaatgt ctatagattg gctttaaaaa ctaccttaaa aagagcaaga aggagtatgg
      601 aacttatagc aagagaggat gaaaatccaa aggttgctga agttatctat ccaataatgc
      661 aggttaatgg ttgtcattat aagggcgttg atgttgcagt tggagggatg gagcagagaa
      721 aaatacacat gttagcaagg gagcttttac caaaaaaggt tgtttgtatt cacaaccctg
      781 tcttaacggg tttggatgga gaaggaaaga tgagttcttc aaaagggaat tttatagctg
      841 ttgatgactc tccagaagag attagggcta agataaagaa agcatactgc ccagctggag
      901 ttgttgaagg aaatccaata atggagatag ctaaatactt ccttgaatat cctttaacca
      961 taaaaaggcc agaaaaattt ggtggagatt tgacagttaa tagctatgag gagttagaga
     1021 gtttatttaa aaataaggaa ttgcatccaa tggatttaaa aaatgctgta gctgaagaac
     1081 ttataaagat tttagagcca attagaaaga gattataagc ggccgcgttt aaacggtctc
     1141 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     1201 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     1261 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     1321 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1381 ggccttgttt gtgagctccc ggtcatcaat catccccata atccttgtta gcctgcaggt
     1441 aattccgctt cgcaacatgt gagcaccggt ttattgacta ccggaagcag tgtgaccgtg
     1501 tgcttctcaa atgcctgagg ccagtttgct caggctctcc ccgtggaggt aataattgac
     1561 gatatgatca gtgcacggct aactaagcgg cctgctgact ttctcgccga tcaaaaggca
     1621 ttttgctatt aagggattga cgagggcgta tctgcgcagt aagatgcgcc ccgcattccg
     1681 gcggtagttc agcagggcag aacggcggac tctaaatccg catggcaggg gttcaaatcc
     1741 cctccgccgg accaaattcg aaaagcctgc tcaacgagca ggcttttttg catgctcgag
     1801 cagctcaggg tcgaatttgc catggcggcc accaggtacc accggcgcct caggcatttg
     1861 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     1921 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg
     1981 cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg gtgatctcgc
     2041 ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt
     2101 cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
     2161 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
     2221 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
     2281 agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat
     2341 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
     2401 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
     2461 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
     2521 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
     2581 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
     2641 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
     2701 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
     2761 ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat
     2821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2881 agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg agactgcggc
     2941 gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag gggactaaca
     3001 tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc gccctcctgc
     3061 cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga ggctcaacca
     3121 tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg tttccggcgg
     3181 gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac ctgttccgcc
     3241 tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta tctcggctcg
     3301 gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca gcccgactgc
     3361 tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg taaaacgcca
     3421 ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta aacacgcggt
     3481 tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg gttgctgtgc
     3541 tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc ttcgatcaaa
     3601 ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag aaaaaaagga
     3661 tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt gcaatttatc
     3721 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     3781 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     3841 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     3901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3961 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     4021 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4081 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4141 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     4201 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4261 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4321 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4381 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4441 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4501 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4561 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4621 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4681 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4741 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4801 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4861 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4921 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattc
//
