LOCUS       62056_4085        2697 bp DNA     circular SYN 16-JUN-2021
DEFINITION  Cloning vector pBla_natural, complete sequence.
ACCESSION   MW816822
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2697)
  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 2697)
  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 2697)
  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..2697
                     /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)"
     CDS             957..1814
                     /label=AmpR
                     /note="beta-lactamase"
     misc_feature    1849..1884
                     /label=proK terminator
                     /note="proK terminator"
     promoter        1914..1944
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    1956..1962
                     /label=5' leader from serT
                     /note="5' leader from serT"
     misc_feature    1963..2024
                     /label=tRNA Ser(UGA)
                     /note="tRNA Ser(UGA)"
     misc_feature    1977..1982
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1983..1988
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1989..1994
                     /label=BsaI
                     /note="BsaI"
     misc_feature    2025..2067
                     /label=3' UTR from serT
                     /note="3' UTR from serT"
     misc_feature    2068..2103
                     /label=proK terminator
                     /note="proK terminator"
     promoter        2151..2198
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2217..2588
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      2603..2697
                     /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 gacaccggta 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 cgccccgaag
     1141 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1201 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1261 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1321 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1381 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1441 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1501 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1561 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1621 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagtcgcg
     1681 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1741 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1801 tgattaagca ttggtaactg tcagaccaag gccaccgctg agcaataaaa ttcgaaaagc
     1861 ctgctcaacg agcaggcttt tttgctgaaa ggaggaacta tatccggatt ggttttacac
     1921 tttatgcttc cggctcgtat gttgcctcta gaaaatcatt ccggaagtgt ggccgagaga
     1981 ccggtaccgg tctctccaga gttcgaatct ctgcgcttcc gccaaataag ataaggggtt
     2041 agctaaatgc taaccccttt ttcttttaat tcgaaaagcc tgctcaacga gcaggctttt
     2101 ttgcctgaac gagcaggctt tgtgcgcgga acccctattt gtttaccgtc gttgacaatt
     2161 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     2221 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     2281 tctggaccga ccggctcggg ttctcccgcg acttcgtgga ggacgacttc gccggtgtgg
     2341 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     2401 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     2461 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     2521 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     2581 agcaggactg agagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     2641 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaat
//
