LOCUS       62056_19385        4839 bp DNA     circular SYN 16-JUN-2021
DEFINITION  Cloning vector pSC101-P_bla_PtNTT2, complete sequence.
ACCESSION   MW816820
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4839)
  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 4839)
  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 4839)
  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..4839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    71..184
                     /label=pUC19-like
                     /note="pUC19-like"
     regulatory      185..235
                     /regulatory_class="promoter"
     misc_feature    235
                     /label=TSS
                     /note="TSS"
     misc_feature    268..273
                     /label=Shine-Dalgarno
                     /note="Shine-Dalgarno"
     CDS             280..1815
                     /codon_start=1
                     /transl_table=11
                     /product="PtNTT2"
                     /label=PtNTT2
                     /protein_id="QWO78774.1"
                     /translation="MGGSTVAPTTPLATGGALRKVRQAVFPIYGNQEVTKFLLIGSIKF
                     FIILALTLTRDTKDTLIVTQCGAEAIAFLKIYGVLPAATAFIALYSKMSNAMGKKMLFY
                     STCIPFFTFFGLFDVFIYPNAERLHPSLEAVQAILPGGAASGGMAVLAKIATHWTSALF
                     YVMAEIYSSVSVGLLFWQFANDVVNVDQAKRFYPLFAQMSGLAPVLAGQYVVRFASKAV
                     NFEASMHRLTAAVTFAGIMICIFYQLSSSYVERTESAKPAADNEQSIKPKKKKPKMSMV
                     ESGKFLASSQYLRLIAMLVLGYGLSINFTEIMWKSLVKKQYPDPLDYQRFMGNFSSAVG
                     LSTCIVIFFGVHVIRLLGWKVGALATPGIMAILALPFFACILLGLDSPARLEIAVIFGT
                     IQSLLSKTSKYALFDPTTQMAYIPLDDESKVKGKAAIDVLGSRIGKSGGSLIQQGLVFV
                     FGNIINAAPVVGVVYYSVLVAWMSAAGRLSGLFQAQTEMDKADKMEAKTNKEK"
     terminator      1849..1943
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1960..2065
                     /label=pCDF-1b-like
                     /note="pCDF-1b-like"
     CDS             complement(2166..2822)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2823..2925)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      3270..3492
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3540..4487
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
ORIGIN
        1 aagctgccga caacacggga gccagtgacg cctcccgtgg ggaaaaaatc atggcaattc
       61 tggaagaaat agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      121 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      181 gtaaattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgcctgt
      241 agaaataatt ttgtttgaat tctaataagg agatatacca tgggaggcag tactgttgca
      301 ccaactacac cgttggcaac cggcggtgcg ctccgcaaag tgcgacaagc cgtctttccc
      361 atctacggaa accaagaagt caccaaattt ctgctcatcg gatccattaa attctttata
      421 atcttggcac tcacgctcac gcgtgatacc aaggacacgt tgattgtcac gcaatgtggt
      481 gccgaagcga ttgcctttct caaaatatac ggggtgctac ccgcagcgac cgcatttatc
      541 gcgctctatt ccaaaatgtc caacgccatg ggcaaaaaaa tgctatttta ttccacttgc
      601 attcctttct ttaccttttt cgggctgttt gatgttttca tttacccgaa cgcggagcga
      661 ctgcacccta gtttggaagc cgtgcaggca attctcccgg gcggtgccgc atctggcggc
      721 atggcggttc tggccaagat tgcgacacac tggacatcgg ccttatttta cgtcatggcg
      781 gaaatatatt cttccgtatc ggtggggcta ttgttttggc agtttgcgaa cgacgtcgtc
      841 aacgtggatc aggccaagcg cttttatcca ttatttgctc aaatgagtgg cctcgctcca
      901 gttttagcgg gccagtatgt ggtacggttt gccagcaaag cggtcaactt tgaggcatcc
      961 atgcatcgac tcacggcggc cgtaacattt gctggtatta tgatttgcat cttttaccaa
     1021 ctcagttcgt catatgtgga gcgaacggaa tcagcaaagc cagcggcaga taacgagcag
     1081 tctatcaaac cgaaaaagaa gaaacccaaa atgtccatgg ttgaatcggg gaaatttctc
     1141 gcgtcaagtc agtacctgcg tctaattgcc atgctggtgc tgggatacgg cctcagtatt
     1201 aactttaccg aaatcatgtg gaaaagcttg gtgaagaaac aatatccaga cccgctagat
     1261 tatcaacgat ttatgggtaa cttctcgtca gcggttggtt tgagcacatg cattgttatt
     1321 ttcttcggtg tgcacgtgat ccgtttgttg gggtggaaag tcggagcgtt ggctacacct
     1381 gggatcatgg ccattctagc gttacccttt tttgcttgca ttttgttggg tttggatagt
     1441 ccagcacgat tggagatcgc cgtaatcttt ggaacaattc agagtttgct gagcaaaacc
     1501 tccaagtatg cccttttcga ccctaccaca caaatggctt atattcctct ggacgacgaa
     1561 tcaaaggtca aaggaaaagc ggcaattgat gttttgggat cgcggattgg caagagtgga
     1621 ggctcactga tccagcaggg cttggtcttt gtttttggaa atatcattaa tgccgcacct
     1681 gtagtagggg ttgtctacta cagtgtcctt gttgcgtgga tgagcgcagc tggccgacta
     1741 agtgggcttt ttcaagcaca aacagaaatg gataaggccg acaaaatgga ggcaaagacc
     1801 aacaaagaaa agtagttaac ctaggctgcg agctccgctg agcaataaga ctcctgttga
     1861 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     1921 gggcgttttt tattggtgag aatttttggc ggatggcatt tgagaagcac acggtcacac
     1981 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     2041 cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt
     2101 attatcactt attcaggcgt agcaccaggc gtttaagggc accaataact gccttaaaaa
     2161 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     2221 acatggaagc catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     2281 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     2341 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     2401 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     2461 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     2521 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     2581 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
     2641 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     2701 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     2761 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     2821 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     2881 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc
     2941 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg
     3001 aagtgatctt ccgtcacagg tatttattcg gcgcctgtag tgccatttac ccccattcac
     3061 tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa agacagcgac tcaggtgcct
     3121 gatggtcgga gacaaaagga atattcagcg atttgcccga gcttgcgagg gtgctactta
     3181 agcctttagg gttttaaggt ctgttttgta gaggagcaaa cagcgtttgc gacatccttt
     3241 tgtaatactg cggaactgac taaagtagtg agttatacac agggctggga tctattcttt
     3301 ttatcttttt ttattctttc tttattctat aaattataac cacttgaata taaacaaaaa
     3361 aaacacacaa aggtctagcg gaatttacag agggtctagc agaatttaca agttttccag
     3421 caaaggtcta gcagaattta cagataccca caactcaaag gaaaaggact agtaattatc
     3481 attgactagc ccatctcaat tggtatagtg attaaaatca cctagaccaa ttgagatgta
     3541 tgtctgaatt agttgttttc aaagcaaatg aactagcgat tagtcgctat gacttaacgg
     3601 agcatgaaac caagctaatt ttatgctgtg tggcactact caaccccacg attgaaaacc
     3661 ctacaaggaa agaacggacg gtatcgttca cttataacca atacgctcag atgatgaaca
     3721 tcagtaggga aaatgcttat ggtgtattag ctaaagcaac cagagagctg atgacgagaa
     3781 ctgtggaaat caggaatcct ttggttaaag gctttgagat tttccagtgg acaaactatg
     3841 ccaagttctc aagcgaaaaa ttagaattag tttttagtga agagatattg ccttatcttt
     3901 tccagttaaa aaaattcata aaatataatc tggaacatgt taagtctttt gaaaacaaat
     3961 actctatgag gatttatgag tggttattaa aagaactaac acaaaagaaa actcacaagg
     4021 caaatataga gattagcctt gatgaattta agttcatgtt aatgcttgaa aataactacc
     4081 atgagtttaa aaggcttaac caatgggttt tgaaaccaat aagtaaagat ttaaacactt
     4141 acagcaatat gaaattggtg gttgataagc gaggccgccc gactgatacg ttgattttcc
     4201 aagttgaact agatagacaa atggatctcg taaccgaact tgagaacaac cagataaaaa
     4261 tgaatggtga caaaatacca acaaccatta catcagattc ctacctacat aacggactaa
     4321 gaaaaacact acacgatgct ttaactgcaa aaattcagct caccagtttt gaggcaaaat
     4381 ttttgagtga catgcaaagt aagtatgatc tcaatggttc gttctcatgg ctcacgcaaa
     4441 aacaacgaac cacactagag aacatactgg ctaaatacgg aaggatctga ggttcttatg
     4501 gctcttgtat ctatcagtga agcatcaaga ctaacaaaca aaagtagaac aactgttcac
     4561 cgttacatat caaagggaaa actgtccata tgcacagatg aaaacggtgt aaaaaagata
     4621 gatacatcag agcttttacg agtttttggt gcattcaaag ctgttcacca tgaacagatc
     4681 gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa caggtgaaac cagtaaaaca
     4741 aagcaactag aacatgaaat tgaacacctg agacaacttg ttacagctca acagtcacac
     4801 atagacagcc tgaaacaggc gatgctgctt atcgaatca
//
