LOCUS       62056_4090        2593 bp DNA     circular SYN 16-JUN-2021
DEFINITION  Cloning vector pBla_T7, complete sequence.
ACCESSION   MW816821
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2593)
  AUTHORS   Hashimoto K, Fischer EC, Romesberg FE.
  TITLE     Efforts toward Further Integration of an Unnatural Base Pair into 
            the Biology of a Semisynthetic Organism
  JOURNAL   J Am Chem Soc (2021) In press
  PUBMED    34096294
REFERENCE   2  (bases 1 to 2593)
  AUTHORS   Hashimoto K.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2021) Department of Chemistry, The Scripps 
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 
            92037, USA
REFERENCE   3  (bases 1 to 2593)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Am Chem 
            Soc (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-MAR-2021) Department of Chemistry, The Scripps Research 
            Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..2593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      109..654
                     /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        807..825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             840..862
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            871..1701
                     /gene="bla"
                     /label=bla
     CDS             join(871..1048,1067..1701)
                     /codon_start=1
                     /transl_table=11
                     /gene="bla"
                     /product="beta-lactamase TEM-1"
                     /label=bla
                     /protein_id="QWO78777.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLDSGKILESFRRAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVREL
                     CSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTT
                     MPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGER
                     GSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     misc_feature    1044..1048
                     /gene="bla"
                     /label=UBP
                     /note="UBP"
     misc_feature    1049..1054
                     /gene="bla"
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1055..1060
                     /gene="bla"
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1061..1066
                     /gene="bla"
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1067..1073
                     /gene="bla"
                     /label=UBP
                     /note="UBP"
     terminator      1733..1780
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1810..1828
                     /note="promoter for bacteriophage T7 RNA polymerase; T7 
                     promoter"
                     /regulatory_class="promoter"
     misc_feature    1840..1846
                     /label=5' leader from serT
                     /note="5' leader from serT"
     misc_feature    1847..1908
                     /label=tRNA Ser(UGA)
                     /note="tRNA Ser(UGA)"
     misc_feature    1861..1866
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1867..1872
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1873..1878
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1909..1951
                     /label=3' UTR from serT
                     /note="3' UTR from serT"
     regulatory      1952..1999
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase; T7 terminator"
                     /regulatory_class="terminator"
     promoter        2047..2094
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2113..2484
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      2499..2593
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 cttaacgtga gttttcgttc cactgaggct gacttcaggt gctacatttg aagagataaa
       61 ttgcactgaa atctagtaat attttatctg attaataaga tgatcttctt gagatcgttt
      121 tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc
      181 gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc
      241 accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc
      301 taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact
      361 caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac
      421 agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa
      481 cgcggccata acagcggaat gacaccggaa aaccgaaagg caggaacagg agagcgcacg
      541 agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac
      601 tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc
      661 tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc
      721 cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag
      781 tgagcgagga agcggaatat atcccttaat acgactcact ataggcctct agaaataatt
      841 ttgtttaact ttaagaagga gatatacatt atgagtattc aacatttccg tgtcgccctt
      901 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      961 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcgac
     1021 agcggtaaga tccttgagag ttttcgccga gaccggtacc ggtctcgcgc ggtattatcc
     1081 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1141 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1201 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1261 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1321 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1381 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1441 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1501 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagt
     1561 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1621 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1681 tcactgatta agcattggta actgtcagac caaggccacc gctgagcaat aactagcata
     1741 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1801 cggattggtt aatacgactc actataggcc tctagaaaat cattccggaa gtgtggccga
     1861 gagaccggta ccggtctctc cagagttcga atctctgcgc ttccgccaaa taagataagg
     1921 ggttagctaa atgctaaccc ctttttcttt tctagcataa ccccttgggg cctctaaacg
     1981 ggtcttgagg ggttttttgc ctgaacgagc aggctttgtg cgcggaaccc ctatttgttt
     2041 accgtcgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     2101 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     2161 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgcgactt cgtggaggac
     2221 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     2281 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     2341 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     2401 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     2461 cacttcgtgg ccgaggagca ggactgagag ctcgcttgga ctcctgttga tagatccagt
     2521 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     2581 tattggtgag aat
//
