LOCUS       40924_35618        6596 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPtPBR2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6596)
  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 6596)
  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 6596)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6596)
  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..6596
                     /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"
     rep_origin      complement(4769..5357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5531..6388)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6389..6493)
                     /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 cactctagag tcgacctgca ggcatctctt ccgcttcctc gctcactgac tcgctgcgct
     4621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5821 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6121 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6541 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaa
//
