LOCUS       40924_35613        7300 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPtPBR11, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7300)
  AUTHORS   Diner RE, Bielinski VA, Dupont CL, Allen AE, Weyman PD.
  TITLE     Refinement of the Diatom Episome Maintenance Sequence and 
            Improvement of Conjugation-Based DNA Delivery Methods
  JOURNAL   Front Bioeng Biotechnol 4, 65 (2016)
  PUBMED    27551676
REFERENCE   2  (bases 1 to 7300)
  AUTHORS   Diner RE, Weyman PD.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JUL-2016) Scripps Institution of Oceanography, 
            University of California San Diego, 9500 Gilman Drive, La Jolla, CA 
            92093, USA
REFERENCE   3  (bases 1 to 7300)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7300)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Bioeng Biotechnol 4, 65 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-JUL-2016) Scripps Institution of Oceanography, University of 
            California San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     regulatory      complement(44..49)
                     /label=P1 promoter
                     /note="P1 promoter"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(64..75)
                     /label=P1 promoter -35 sequence region
                     /note="P1 promoter -35 sequence region"
                     /regulatory_class="minus_35_signal"
     CDS             86..1273
                     /label=TcR
                     /note="tetracycline efflux protein"
     regulatory      1905..1909
                     /label=rop RBS
                     /note="rop RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1915..2103
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(2340..2708)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(2741..2850)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     primer_bind     3127..3143
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3167..3185
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3889..4260
                     /label=BleoR
                     /note="antibiotic-binding protein"
     misc_feature    4563..5066
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        5067..5253
                     /label=HIS3 promoter
     rep_origin      complement(5473..6061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6235..7092)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7093..7197)
                     /label=AmpR promoter
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
      421 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
      481 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
      541 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
      601 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
      661 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
      721 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
      781 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
      841 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
      901 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
      961 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     1021 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     1081 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc
     1141 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     1201 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     1261 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     1321 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     1381 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
     1441 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     1501 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     1561 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     1621 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     1681 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     1741 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     1801 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     1861 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     1921 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     1981 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     2041 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     2101 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     2161 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgg atcgtcttgc
     2221 cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca
     2281 tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat
     2341 ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct
     2401 tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg
     2461 tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc
     2521 agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc
     2581 agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt
     2641 tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca
     2701 tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca
     2761 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     2821 gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag
     2881 tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa
     2941 aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aagatgccat tcgccattca
     3001 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3061 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3121 gacgttgtaa aacgacggcc agtgaccatg attacgccaa gctcgaaatt aaccctcact
     3181 aaagggaaca aaagctggta cctaacagga ttagtgcaat tcgagttgaa tcactgggaa
     3241 aaacattgtc ttctttttta tattatcatt tgcattagtg ctgcagtcgt agatacttgt
     3301 tggttgaaag acatcagctg ggagggactg gactagcgtt tggtaaggag acatacctgt
     3361 taacgttggt tgcaaaattc catttcgcga tttatgttat ctgtaaatcc tgatttgtct
     3421 ggaattcttg atacttccgt ttttttagag gccaatgatt agcatcggcg attctcaaaa
     3481 tagcattttc gacatgcggt gctgatttca taaacataga caacgctttt acatgtaaaa
     3541 gtaacttgcg gacttggaac agtgctctgt ttttggtgtg aacgtaactc agcaatattt
     3601 ctgtgctagc aaggtttttt atgatcgacc gaagatctca aaactccggg tctttcaact
     3661 gtctgactag accatgttcg taacgtcgga cagcagcttt cgttgtactg gtagaatttc
     3721 tacgtgcgaa gcacgtgtag gcaggttgaa cgacgatccc tgccgatgga tggattggca
     3781 cgcggcggaa cgctttcgtg atctacacca cctggatctt cacatatctt cgaaatcgaa
     3841 aaattaacca agtcgacggt atcgataata ttctagctga gggtacccat ggccaagttg
     3901 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     3961 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     4021 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     4081 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     4141 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     4201 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     4261 tgaccgacgc cgaccaacac cgccggtccg acgcggcccg acgggtccga ggcctcggag
     4321 atctgggccc atgcggccgc aacaactacc tcgactttgg ctgggacact ttcagtgagg
     4381 acaagaagct tcagaagcgt gctatcgaac tcaaccaggg acgtgcggca caaatgggca
     4441 tccttgctct catggtgcac gaacagttgg gagtctctat ccttccttaa aaatttaatt
     4501 ttcattagtt gcagtcactc cgctttggtt tcacagtcag gaataacact agctcgtctt
     4561 caatcacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
     4621 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
     4681 agactctagg gggatcgcca acaaatacta ccttttacct tgctcttcct gctctcaggt
     4741 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
     4801 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
     4861 taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt
     4921 ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac
     4981 ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg
     5041 cgcgtgtaag ttacaggcaa gcgatcctag tacactctat atttttttat gcctcggtaa
     5101 tgattttcat tttttttttc cacctagcgg atgactcttt ttttttctta gcgattggca
     5161 ttatcacata atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta
     5221 aaaaatgagc aggcaagata aacgaaggca aagatgacag agcagactct agagtcgacc
     5281 tgcaggcatc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5341 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5401 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5641 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5881 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6301 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6361 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6421 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6481 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6541 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6601 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6661 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6721 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6781 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6841 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7081 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7141 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7201 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7261 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
//
