LOCUS       sequence-111717-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Episome vector for Thalassiosira pseudonana (pUC-based).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Karas BJ, Diner RE, Lefebvre SC, McQuaid J, Phillips AP, Noddings 
            CM, Brunson JK, Valas RE, Deerinck TJ, Jablanovic J, Gillard JT, 
            Beeri K, Ellisman MH, Glass JI, Hutchison CA 3rd, Smith HO, Venter 
            JC, Allen AE, Dupont CL, Weyman PD
  TITLE     Designer diatom episomes delivered by bacterial conjugation.
  JOURNAL   Nat Commun. 2015 Apr 21;6:6925. doi: 10.1038/ncomms7925.
  PUBMED    25897682
REFERENCE   2  (bases 1 to 7793)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2015-04-21"; volume: "6:6925. doi"; pages: " 
            10.1038/ncomms7925"
COMMENT     Sequence Label: pTpPuc3
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(307..678)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            711..820
                     /note="incP origin of transfer"
     primer_bind     1083..1105
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1097..1114
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1098..1114
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2096..2665
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     misc_feature    3183..3686
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3687..3873
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3874..4533
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     primer_bind     complement(4602..4618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4602..4618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4615..4637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4626..4642
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4650..4656,4657..4674,4675..4680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4695..4716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4833..4850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5004..5592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5084..5103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6429..6448)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6440..7255
                     /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             6440..7255
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6513..6532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7356..7375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7594..7698)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7766..7784
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accagatcgt cttgccttgc tcgtcggtga tgtacttaca gctcgaagtg cctcttcttg
      241 atggagcgca tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta
      301 aaaaagtcat ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc
      361 ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt
      421 gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt
      481 gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg
      541 gccgacggcc agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat
      601 cgctcttcgt tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg
      661 cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc
      721 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
      781 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
      841 ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg
      901 atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aagatggcca
      961 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     1021 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     1081 ttcccagtca cgacgttgta aaacgacggc cagtgagctt gcgctttttc cgagaactcc
     1141 ccataagtca acggctccaa tcaagaatgt atccgacaac ggcgagcata gcaacacgtc
     1201 cgtctttgga gtagaatcat catgttgtgg atgaatacac agatgaatga cattaaaagc
     1261 atgaacatgt tagagagtag gaggtagaga ttgatatggt agcattgcga tgtttgtttt
     1321 tggtcagcat atgatgagtg gataccaata tgatgaaagt tgaatctcgc gtttgagctc
     1381 agcggtacgt tattgatcga aagtagcctg atcaaaatcc ttggagagta caagaggatc
     1441 aaagaatcca gtgggggcga taactccaag ctcgttctca aagaggcaat ggaggtagaa
     1501 actcatccca gttgagaaga agtgaaggca gtggcggtgg cgaaagcaga ggcaacgagg
     1561 acagacttcc tgtgggttga tgcaacgaat atttccagaa ggagaagttt agagagttga
     1621 accgctacct acaatgacaa agtatcgtat cgattttgat gttggttggt tatgaattca
     1681 aactgtaagt tggattgtga gaagatcaga agttgaacga acacatcttt ccgatcattc
     1741 acctccacac tgcaacaaca cggtacttct tccgcggcag gtctctgtcg ccattctctt
     1801 gtcctgttgt tggctgtgag acgaggaaag caacgacaag tttcacaaaa gggagttcct
     1861 ttaacgagat atgtttttta taaagagtcc caatagaaag acaaattgat tcctccgtgc
     1921 aaacgcgcaa ataaacacca cgtccattat atccatatct ttcagagtat ccaacaagtg
     1981 ttgaaggaca ggtagttgaa gtaacgtatc ttccccctcg actggatcca tcaacaaggc
     2041 gaacaaatcc attcaacctc tcataaatta tctgatttac caaaccgata ccaaaatgac
     2101 cactcttgac gacacggctt accggtaccg caccagtgtc ccgggggacg ccgaggccat
     2161 cgaggcactg gatgggtcct tcaccaccga caccgtcttc cgcgtcaccg ccaccgggga
     2221 cggcttcacc ctgcgggagg tgccggtgga cccgcccctg accaaggtgt tccccgacga
     2281 cgaatcggac gacgaatcgg acgacgggga ggacggcgac ccggactccc ggacgttcgt
     2341 cgcgtacggg gacgacggcg acctggcggg cttcgtggtc gtctcgtact ccggctggaa
     2401 ccgccggctg accgtcgagg acatcgaggt cgccccggag caccgggggc acggggtcgg
     2461 gcgcgcgttg atggggctcg cgacggagtt cgcccgcgag cggggcgccg ggcacctctg
     2521 gctggaggtc accaacgtca acgcaccggc gatccacgcg taccggcgga tggggttcac
     2581 cctctgcggc ctggacaccg ccctgtacga cggcaccgcc tcggacggcg agcaggcgct
     2641 ctacatgagc atgccctgcc cctgagcggc cgcatactgg attggtgaat caatgagccg
     2701 tagcacaatg gttacattcg gctagctaag atccaatggc aaggaccaag tgctggaact
     2761 tgttttgctt tagcagatct tagcgtgaga ggtatttgtc ctctgtcagg agtagatagt
     2821 agatgttctt tttaaactaa aatgctaact gttccgaatt cctcatcgca gctaatccgt
     2881 acatcaaaag acaaaatgct aggtatgtgt actacatctc ctgttgctag ataagacata
     2941 tgataggaaa cacaccatca atagtcattg tagctttact tatactacgc atttgcactt
     3001 tcccctgagt ggcagaggcg cattgagaaa atcgatctca acatagttta tgtagcatcc
     3061 cctagatcca ttactttaag tctccttcgt ctttggtgta ggcatgttgg acacaacgag
     3121 gtaaaacaca acacaaacaa tgtgtccagc aaagtagtag ctgctccagt tctcccgcga
     3181 gcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
     3241 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
     3301 agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct gctctcaggt
     3361 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
     3421 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
     3481 taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt
     3541 ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac
     3601 ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg
     3661 cgcgtgtaag ttacaggcaa gcgatcctag tacactctat atttttttat gcctcggtaa
     3721 tgattttcat tttttttttc cacctagcgg atgactcttt ttttttctta gcgattggca
     3781 ttatcacata atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta
     3841 aaaaatgagc aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag
     3901 cgtattacaa atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg
     3961 atagagcact cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa
     4021 tcgcaagtga ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg
     4081 gccaagcatt ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac
     4141 catcacacca ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg
     4201 gcgcgtggag taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga
     4261 gcggtggtag atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag
     4321 aaagtaggag atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct
     4381 agcagaatta ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag
     4441 agtgcgttca aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac
     4501 gatgttccct ccaccaaagg tgttcttatg tagttttaca caggagtctg gacttgacct
     4561 ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4621 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4681 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4741 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6301 tctgtgactg gtgagtcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg
     6361 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     6421 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct cgaggccgcg
     6481 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     6541 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     6601 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     6661 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     6721 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     6781 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     6841 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     6901 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     6961 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     7021 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     7081 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     7141 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     7201 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     7261 attggttaat tggttgtaac actggcactc aaccaagtca ttctgagaat agtgtatgcg
     7321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7621 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7681 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7741 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
