LOCUS       C0.394_trcE-star        3471 bp DNA     circular SYN 06-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3471)
  AUTHORS   SynthSys Center
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3471
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     terminator      78..172
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      328..357
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    365..368
                     /label=GGAG
     promoter        369..423
                     /label=trc promoter
     misc_feature    424..466
                     /label=Leader sequence
                     /note="Se Atsumi paper Figure 3 (Ptrc Leader Sequence)"
     misc_feature    424
                     /label=TSS
     misc_signal     467..502
                     /label=Riboswith E* aptamer
     RBS             510..516
                     /label=RBS
                     /note="In prokaryotes (such as E. coli) the RBS typically
                     lies about 7 nucleotides upstream from the start codon 
                     (i.e., the first AUG). The sequence itself in general is 
                     called the ""Shine-Dalgarno"" sequence after its 
                     discoverers, regardless of the exact identity of the bases.
                     Strong Shine-Dalgarno sequences are rich in purines 
                     (A's,G's), and the ""Shine-Dalgarno consensus"" sequence --
                     derived statistically from lining up many 
                     well-characterized strong ribosome binding sites -- has the
                     sequence AGGAGG. The complementary sequence (CCUCCU) occurs
                     at the 3'-end of the structural RNA (""16S"") of the small 
                     ribosomal subunit and it base-pairs with the Shine-Dalgarno
                     sequence in the mRNA to facilitate proper initiation of 
                     protein synthesis."
     misc_feature    523..526
                     /label=AATG
     CDS             complement(664..1479)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1837..2787)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with
                     thepSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2835..3057
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta
       61 ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
      121 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
      181 ctcgtaaact tggtctgaca gctctagctc cggcaaaaaa acgggcaagg tgtcaccacc
      241 ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt tgccacctga cgtctaagaa
      301 aaggaatatt cagcaatttg cccgtgccga agaaaggccc acccgtgaag gtgagccggt
      361 ctcaggagaa atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg
      421 tggaattgtg agcggataac aatttcatac gctcacaatt ggtaccggtg ataccagcat
      481 cgtcttgatg cccttggcag caccctgcta aggaggcaac agaatgtgag accagtcaaa
      541 agcctccgac cggaggcttt tgactaaaac ttcccttggg gttatcattg gggctcactc
      601 aaaggcggta atcagataaa aaaaatcctt agctttcgct aaggatgatt tctgctagta
      661 ttattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
      721 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      781 ccaaagaatg gcaaggtcct ggtaacggtc tgcgattccg acccgtccaa catcaataca
      841 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      901 gactgaatcc ggtgagaatg gcaagagctt gtgcatttct ttccagactt gttcaacagg
      961 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcatgcgtga
     1021 ttgcgcctga gcaagacgaa atacacgatc gctgttaaaa ggacaattac aaacaggaat
     1081 cgaatgtaac cggcgcagga acacggccag cgcatcaaca atattttcac ctgaatcagg
     1141 atattcttct aatacctgga aggctgtttt cccaggaatc gcggtggtga gtaaccacgc
     1201 atcatcagga gtacggataa aatgcttgat ggtcgggaga ggcataaact ccgtcagcca
     1261 gttgagacgg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     1321 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc
     1381 gacattatcg cgagcccatt tatacccata taaatcagcg tccatgttgg agtttaagcg
     1441 cggacgggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     1501 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     1561 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     1621 ggatcagctc tagtagttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
     1681 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     1741 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     1801 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     1861 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     1921 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     1981 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     2041 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     2101 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     2161 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     2221 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     2281 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     2341 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     2401 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     2461 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     2521 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     2581 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     2641 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     2701 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     2761 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     2821 ataccaattg agatgggcta gtcaatgata attacatgtc cttttccttt gagttgtggg
     2881 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     2941 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     3001 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     3061 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     3121 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     3181 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     3241 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     3301 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     3361 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
     3421 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc a
//
