LOCUS       sequence-343816-        6451 bp    DNA     circular SYN 15-APR-2025
DEFINITION  bacterial co-expression of Drosophila Sep2 and of Drosophila Peanut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6451)
  AUTHORS   Iv F, Martins CS, Castro-Linares G, Taveneau C, Barbier P, 
            Verdier-Pinard P, Camoin L, Audebert S, Tsai FC, Ramond L, Llewellyn
            A, Belhabib M, Nakazawa K, Di Cicco A, Vincentelli R, Wenger J, 
            Cabantous S, Koenderink GH, Bertin A, Mavrakis M
  TITLE     Insights into animal septins using recombinant human septin octamers
            with distinct SEPT9 isoforms.
  JOURNAL   J Cell Sci. 2021 Jul 8. pii: 270842. doi: 10.1242/jcs.258484.
  PUBMED    34237139
REFERENCE   2  (bases 1 to 6451)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell Sci.
            2021 Jul 8. pii: 270842. doi: 10.1242/jcs.258484."
COMMENT     Sequence Label: pnCS_DSep2_Peanut-TEV-Strep
FEATURES             Location/Qualifiers
     source          1..6451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     81..100
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        81..99
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    100..124
                     /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)."
     RBS             139..161
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1453..1475
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3100..3120
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3133..3156
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3199..3217)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3213..3260
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3431..4222)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4223..4314)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4362..5100)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5310..6392)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6354..6373
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(6393..6451,1..19))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
       61 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
      121 ttccccacta gtaataattt tgtttaactt taagaaggag atatacatat gtccgttgag
      181 gtggatttcg tcgacaagaa ggaggtgcac ctccgcaccc tgaagcagtc cggtcacgtg
      241 ggattcgaca gcttgcctga ccaactggtc aacaagagtg tccagaatgg attcgtcttc
      301 aacgtcatgt gtataggcga aaccggactg ggcaagtcga cgctgatgga cacgctgttc
      361 aacaccagct tcgagtccac gccgagtccc cacactttgc cgagcgtcaa gttgaaggca
      421 cacacctacg agctgcagga gagcaatgtg cgactcaagc tcaccatctg cgacacggtt
      481 ggctacggcg atcagatcaa caaggacgac tcgttcaagg cggtggtgga ctacatcgac
      541 gcacagttcg agaactatct gcaggaggag ctgaagatca agcgatcgct ggtgacatgc
      601 cacgacagcc gcatccacat ctgcctctac tttatttgcc ccactgggca cggcctcaaa
      661 tcgctggacc tggtgtgcat gaagaagctg gacagcaagg tgaacataat tccggtgatt
      721 gccaaggcgg ataccatttc caaggttgag ctacagcgct tcaaggcgaa gatcattcag
      781 gagctaaacg ccaatggagt gcacatctac cagttcccca ccgacgatga gacggtggcc
      841 gagaccaaca ccagcatgaa ctcgcacatt cccttcgccg tggtgggcag cacggagttt
      901 attaaggtgg gcaacaagct catccgcgct cgccagtacc cctggggcac ggtgcaggtg
      961 gagaacgaga cgcactgcga ctttgtgaag ctgcgcgaga tgctcattcg caccaacatg
     1021 gaggatatgc gcgaaaagac acacacgcgc cactacgagc tgtatcgcca gaagaggctg
     1081 gagcaaatgg gcttcagcga tgtggacagc gacaacaagc ccatctcgtt ccagcagacg
     1141 ttcgaggcaa agcgctccaa ccatttggcc gagctgcagt ccaaggagga ggaggttcgt
     1201 cagatgttcg tgcagcgcgt taaggagaag gaggccgagc tgaaggagag cgagaaggac
     1261 ctgcacgcca agttcgagaa gctaaagcgg gatcacgccg aggagaagcg caagctggaa
     1321 gagtcgcgca aggcgctcga ggaggactac cttgacttcc agcggcgcaa gcagcagttg
     1381 gccaccgccc accacacgct caccctcggc aagagcaaga agaagtaggg atcctaatag
     1441 tctagtaata attttgttta actttaagaa ggagatatac atatgaatag tcctcgctcg
     1501 aacgcggtta atggcggcag cggcggcgcc atctccgctc tgcccagcac tctggcccaa
     1561 ctggctctgc gcgacaagca gcaggctgcg tcggcatcgg cgtcctctgc cacgaacggc
     1621 agcagcggca gcgagtcctt ggtgggcgtc ggtggacgac cgcccaatca gccgcccagt
     1681 gtgccagtgg ccgccagtgg caagctggac accagcggcg gaggcgcctc caacggcgac
     1741 tcgaacaaac tgacccacga tctgcaggag aaggagcacc agcaggcgca gaagccacag
     1801 aagccaccgc tcccggtgcg ccagaagccc atggagatcg ccggctatgt gggcttcgcc
     1861 aacctgccca atcaggtcta tcgcaaggcc gtcaagcggg gattcgagtt cactctgatg
     1921 gtggtgggcg ccagtgggct gggcaagtcg acgctgatca actccatgtt cctgtcggac
     1981 atctacaacg cggaacagta tccgggcccc tcgctgcgca aaaagaagac cgtggccgtg
     2041 gaggccacca aagtaatgct caaggagaac ggcgtcaatc tgacactcac ggtggtggac
     2101 acacccggat tcggtgatgc cgttgacaac agcaactgct gggtgcccat actggagtac
     2161 gtggatagca agtacgagga gtacctgaca gccgagtcgc gggtgtaccg aaaaaccatt
     2221 tcggacagcc gggtgcactg ctgcctctac ttcatagcgc catcgggaca cggactcctg
     2281 ccgctggaca ttgcctgcat gcagagcctg tcggacaagg taaatctggt gccggtgatc
     2341 gccaaggcgg acaccatgac gcccgacgag gtgcatctat ttaagaagca gatccttaac
     2401 gagatcgccc agcacaagat caagatctac gacttccccg ccacgctgga ggatgcggcc
     2461 gaggaggcca aaaccacgca gaatttgcgc agccgagtgc cattcgcggt ggtgggtgcc
     2521 aacaccatca tcgagcagga cggaaagaag gtccggggta ggcgttatcc ctggggcctg
     2581 gtggaggtgg agaacctgac gcattgcgac tttatagcgc tgcgcaacat ggtcatacgc
     2641 acgcaccttc aggacctcaa ggacgtgacg aacaacgtcc actacgagaa ctaccgctgc
     2701 cggaagctat ccgagctggg attggtggac ggcaaggcca ggctgtccaa caagaaccca
     2761 ctcacccaga tggaggagga aaagcgggag cacgagcaga agatgaagaa gatggaggcc
     2821 gagatggagc aggtgtttga catgaaggtc aaggagaaga tgcagaagct cagggactcg
     2881 gagctggagc tggcgcggcg ccacgaggag cgcaagaagg cgctggagct gcagatccgg
     2941 gagctggagg agaagagacg cgagttcgag cgcgagaaga aggagtggga ggatgtcaac
     3001 cacgtgacgc tcgaggagct caagcgacgc agcctggggg ccaacagcag cacggacaat
     3061 gtggatggca agaaggagaa gaaaaagaag ggtctgttcg aaaaccttta cttccagggc
     3121 agcggcagcg gctggagcca cccgcagttc gaaaaataag gatcctaata gtctagagct
     3181 agccctaggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3241 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc
     3301 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     3361 gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa
     3421 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt
     3481 cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc
     3541 tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag
     3601 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
     3661 gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt
     3721 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
     3781 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt
     3841 cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt
     3901 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
     3961 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
     4021 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
     4081 tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca
     4141 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
     4201 cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg
     4261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta
     4321 ggtcactcgg tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa
     4381 agttacccac agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc
     4441 cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg
     4501 cttttccggt gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga
     4561 aacccgacag gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc
     4621 ctgttccgac cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt
     4681 ggcgctttct catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgta agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt
     4801 tcacttgagt ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac
     4861 tgggagttcg cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa
     4921 agtccggcta cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac
     4981 ggttaagcag ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt
     5041 tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5101 ttttctactg aaccgctctt gatttcagtg caatttatct cttcaaatgt agcacctgaa
     5161 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
     5221 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5281 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5341 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5401 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5461 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5521 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5581 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5641 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5701 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5761 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5821 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5881 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5941 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     6001 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6061 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6121 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6181 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6241 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6301 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6361 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6421 gggcgctatc atgccatacc gcgaaaggtt t
//
