LOCUS       62056_20335        3165 bp DNA     circular SYN 19-APR-2020
DEFINITION  Cloning vector pSyn_Mz_sfGFP_190(ochre), complete sequence.
ACCESSION   MN882182
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3165)
  AUTHORS   Fischer EC, Hashimoto K, Zhang Y, Feldman AW, Dien VT, Karadeema RJ,
            Adhikary R, Ledbetter MP, Krishnamurthy R, Romesberg FE.
  TITLE     New codons for efficient production of unnatural proteins in a 
            semi-synthetic organism
  JOURNAL   Nat. Chem. Biol. (2020) In press
REFERENCE   2  (bases 1 to 3165)
  AUTHORS   Fischer EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-DEC-2019) Department of Chemistry, The Scripps 
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 
            92037, USA
REFERENCE   3  (bases 1 to 3165)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol. (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-DEC-2019) Department of Chemistry, The Scripps Research 
            Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA"
FEATURES             Location/Qualifiers
     source          1..3165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      48..593
                     /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."
     promoter        746..764
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    766..784
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             799..821
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1552..1575
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             1612..1635
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      1686..1733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1763..1781
                     /note="promoter for bacteriophage T7 RNA polymerase; T7 
                     promoter"
                     /regulatory_class="promoter"
     protein_bind    1782..1806
                     /label=lac operator
                     /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)."
     misc_feature    1818..1822
                     /label=5' proK leader
                     /note="5' proK leader"
     misc_feature    1823..1894
                     /note="tRNA-Pyl; Methanosarcina mazei tRNAPyl(TTA)"
     misc_feature    1895..1933
                     /label=3' proK trailer
                     /note="3' proK trailer"
     regulatory      1934..1981
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase; T7 terminator"
                     /regulatory_class="terminator"
     regulatory      2019..2125
                     /label=proK promoter
                     /note="proK promoter"
                     /regulatory_class="promoter"
     misc_RNA        2146..2221
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      2229..2264
                     /label=proK terminator
                     /note="proK terminator"
                     /regulatory_class="terminator"
     regulatory      2277..2382
                     /label=proK promoter
                     /note="proK promoter"
                     /regulatory_class="promoter"
     misc_RNA        2401..2472
                     /note="guide RNA scaffold for the CRISPR/Cas9 system; gRNA 
                     scaffold"
     regulatory      2480..2515
                     /label=proK terminator
                     /note="proK terminator"
                     /regulatory_class="terminator"
     promoter        2549..2596
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2615..2986
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      3001..3095
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tgcactgaaa tctagtaata ttttatctga ttaataagat gatcttcttg agatcgtttt
       61 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
      121 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
      181 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
      241 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
      301 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
      361 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
      421 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
      481 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
      541 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
      601 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
      661 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
      721 gagcgaggaa gcggaatata tcccttaata cgactcacta tagggtccct atcagtgata
      781 gagaggtcta gaaataattt tgtttaactt taagaaggag atatacatat gtcgaaaggc
      841 gaagaactgt ttacgggagt ggtgcctatc ctggtagagc tcgacggaga tgtaaacggt
      901 cacaaatttt cagtccgcgg ggaaggcgaa ggcgatgcga ccaacggtaa attaactttg
      961 aagtttattt gcaccaccgg caaattaccg gtgccttggc cgacgcttgt gacgaccctg
     1021 acttacgggg tgcagtgttt cagtcgctac ccagatcaca tgaaacgcca tgacttcttc
     1081 aaatctgcga tgccggaagg ctatgtgcag gaacgtacaa ttagctttaa agacgacggc
     1141 acgtataaaa cgcgggcaga ggttaaattt gagggagata ccctggtaaa ccgtattgaa
     1201 ctgaaaggca tcgattttaa agaagatggg aacatcttgg gccacaaact cgagtacaac
     1261 tttaactcac acaatgtata catcacggca gacaaacaaa agaatggaat caaagcaaat
     1321 ttcaagatcc gtcataacgt ggaggacggt tccgtgcagc ttgcagatca ctatcagcag
     1381 aatacgccga ttggctaagg cccggtgctg ctgcccgata atcactacct ctctactcag
     1441 agtgttttat cgaaagaccc gaacgagaag cgtgatcaca tggtgctgct tgaatttgtt
     1501 accgcggcag gtattacaca cggcatggat gagttgtata agggatccgc ttggagccac
     1561 ccgcagttcg agaaaggtgg aggttccgga ggtggatcgg gaggttcggc gtggagccac
     1621 ccgcagttcg aaaaatgaaa gcttaattag ctgagcttgg actccctgcc accgctgagc
     1681 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1741 gaggaactat atccggattg gttaatacga ctcactatag gggaattgtg agcggataac
     1801 aattcccctc tagaaaagca ttggaaacct gatcatgtag atcgaacgga ctttaaatcc
     1861 gttcagccgg gttagattcc cggggtttcc gccaaattcg aaaagcctgc tcaacgagca
     1921 ggcttttttg catctagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1981 gcctgaacga gcaggctttt ttgcataagc ttcctagtgg cagcggctaa ctaagcggcc
     2041 tgctgacttt ctcgccgatc aaaaggcatt ttgctattaa gggattgacg agggcgtatc
     2101 tgcgcagtaa gatgcgcccc gcattggaga cgccatggcg tctcggtttt agagctagaa
     2161 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     2221 ctttttttaa ttcgaaaagc ctgctcaacg agcaggcttt tttggtcgac agtagtggca
     2281 gcggctaact aagcggcctg ctgactttct cgccgatcaa aaggcatttt gctattaagg
     2341 gattgacgag ggcgtatctg cgcagtaaga tgcgccccgc atgagacggc atgccgtctc
     2401 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     2461 ccgagtcggt gcttttttta attcgaaaag cctgctcaac gagcaggctt ttttggtcga
     2521 cagttcatag gtgattgcgg atcccgtcgt tgacaattaa tcatcggcat agtatatcgg
     2581 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     2641 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     2701 ctcccgcgac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     2761 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     2821 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     2881 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     2941 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgag agctcgcttg
     3001 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     3061 tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagctag taacaactta
     3121 tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaat
//
