LOCUS       sequence-174621-        3742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBR322-based entry vector for Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3742)
  AUTHORS   Bielinski VA, Bolt TM, Dupont CL, Weyman PD
  TITLE     Episomal tools for RNAi in the diatom Phaeodactylum tricornutum
  JOURNAL   PeerJ Preprints. 2017 5:e2907v1
REFERENCE   2  (bases 1 to 3742)
  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: "PeerJ 
            Preprints. 2017 5:e2907v1"
COMMENT     Sequence Label: pPtGG-1
FEATURES             Location/Qualifiers
     source          1..3742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..83)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             1541..1732
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1726..1748)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1834..1974
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1889..1908
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1989..2006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2160..2748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2240..2259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2831..2853
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2845..2862
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2846..2862
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2878..2977
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3195..3294)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3378..3383,3384..3400,3401..3406)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             join(3454..3742,1..902)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
        1 ggatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
       61 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
      121 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
      181 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
      241 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg accgatgccc ttgagagcct
      301 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
      361 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
      421 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
      481 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
      541 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
      601 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
      661 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
      721 aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg
      781 cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc
      841 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
      901 gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca
      961 attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc
     1021 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt
     1081 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     1141 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     1201 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     1261 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     1321 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     1381 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     1441 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     1501 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     1561 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     1621 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     1681 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     1741 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     1801 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     1861 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1921 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     1981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2581 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg cgcaacgttg
     2821 ttgccattgt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa
     2881 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc
     2941 ttttttataa tgccaacttt gtacaaaaaa gcaggctccg cggccgcccc cttcacctgg
     3001 gaattcgtta acagatctga gaccgcggct cgagactagt gcctaggtga gtttctattc
     3061 gcagtcggct gatctgtgtg aaatagggat aacagggtaa ttcttaatga agggtccaat
     3121 taccaatttg aaactcagct agctctagaa tatcaattgg gatccggtct catcgacaaa
     3181 gggtgggcgc gccgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3241 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     3301 agctgatatc ccctatagtg aagggtcagt gctgcaacat tatcacgagg ccctttcgtc
     3361 ttcaagaatt ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca
     3421 cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc
     3481 atcctcggca ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg
     3541 ggcctcttgc gggatatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta
     3601 gcgctatatg cgttgatgca atttctatgc gcacccgttc tcggagcact gtccgaccgc
     3661 tttggccgcc gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc
     3721 atggcgacca cacccgtcct gt
//
