LOCUS       62056_4080        2667 bp DNA     circular SYN 16-JUN-2021
DEFINITION  Cloning vector pBla_gg, complete sequence.
ACCESSION   MW816823
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2667)
  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 2667)
  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 2667)
  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..2667
                     /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..911
                     /label=AmpR promoter
     RBS             926..948
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            957..1787
                     /gene="bla"
                     /label=bla
     CDS             join(957..1134,1153..1787)
                     /codon_start=1
                     /transl_table=11
                     /gene="bla"
                     /product="beta-lactamase TEM-1"
                     /label=bla
                     /protein_id="QWO78781.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLDSGKILESFRRAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVREL
                     CSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTT
                     MPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGER
                     GSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     misc_feature    1130..1134
                     /gene="bla"
                     /label=UBP
                     /note="UBP"
     misc_feature    1135..1140
                     /gene="bla"
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1141..1146
                     /gene="bla"
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1147..1152
                     /gene="bla"
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1153..1159
                     /gene="bla"
                     /label=UBP
                     /note="UBP"
     misc_feature    1819..1854
                     /label=proK terminator
                     /note="proK terminator"
     promoter        1884..1914
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    1926..1932
                     /label=5' leader from serT
                     /note="5' leader from serT"
     misc_feature    1933..1994
                     /label=tRNA Ser(UGA)
                     /note="tRNA Ser(UGA)"
     misc_feature    1947..1952
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1953..1958
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1959..1964
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1995..2037
                     /label=3' UTR from serT
                     /note="3' UTR from serT"
     misc_feature    2038..2073
                     /label=proK terminator
                     /note="proK terminator"
     promoter        2121..2168
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2187..2558
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      2573..2667
                     /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 atccctcgcg gaacccctat ttgtttattt ttctaaatac
      841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      901 aaaggaagag tcctctagaa ataattttgt ttaactttaa gaaggagata tacattatga
      961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1021 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1081 tgggttacat cgaactggat ctcgacagcg gtaagatcct tgagagtttt cgccgagacc
     1141 ggtaccggtc tcgcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1201 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1261 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1321 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1381 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1441 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     1501 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1561 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1621 aatctggagc cggtgagcgt ggaagtcgcg gtatcattgc agcactgggg ccagatggta
     1681 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1741 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1801 gccaccgctg agcaataaaa ttcgaaaagc ctgctcaacg agcaggcttt tttgctgaaa
     1861 ggaggaacta tatccggatt ggttttacac tttatgcttc cggctcgtat gttgcctcta
     1921 gaaaatcatt ccggaagtgt ggccgagaga ccggtaccgg tctctccaga gttcgaatct
     1981 ctgcgcttcc gccaaataag ataaggggtt agctaaatgc taaccccttt ttcttttaat
     2041 tcgaaaagcc tgctcaacga gcaggctttt ttgcctgaac gagcaggctt tgtgcgcgga
     2101 acccctattt gtttaccgtc gttgacaatt aatcatcggc atagtatatc ggcatagtat
     2161 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     2221 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccgcg
     2281 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     2341 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     2401 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     2461 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     2521 ccggcaactg cgtgcacttc gtggccgagg agcaggactg agagctcgct tggactcctg
     2581 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     2641 cgccgggcgt tttttattgg tgagaat
//
