LOCUS       sequence-358020-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial co-expression of human SEPT7 and of human SEPT9_i1-i5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  AUTHORS   Kuzmic M, Linares GC, Fialova JL, Iv F, Salaun D, Llewellyn A, Gomes
            M, Belhabib M, Liu Y, Asano K, Rodrigues M, Isnardon D, Tachibana T,
            Koenderink GH, Badache A, Mavrakis M, Verdier-Pinard P
  TITLE     Septin-microtubule association via a motif unique to the isoform 1 
            of septin 9 tunes stress fibers.
  JOURNAL   J Cell Sci. 2021 Dec 2. pii: 273604. doi: 10.1242/jcs.258850.
  PUBMED    34854883
REFERENCE   2  (bases 1 to 6018)
  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: "J Cell Sci.
            2021 Dec 2. pii: 273604. doi: 10.1242/jcs.258850."
COMMENT     Sequence Label: pnCS_SEPT7_SEPT9_i1-i5-TEV-Strep
FEATURES             Location/Qualifiers
     source          1..6018
                     /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)"
     CDS             1246..1266
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1279..1302
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             1318..1340
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2701..2719)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2715..2762
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2933..3724)
                     /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(3725..3816)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4877..5959)
                     /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     5921..5940
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(5960..6018,1..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 gaagaagtct
      181 tactcaggag gcacgcggac ctccagtggc cggctccgga ggcttggtga ctccagtggc
      241 ccagccgaca cccccagaga tgccgggctc aagcaggcgc ctgcatcacg gaacgagaag
      301 gccccggtgg acttcggcta cgtggggatt gactccatcc tggagcagat gcgccggaag
      361 gccatgaagc agggcttcga gttcaacatc atggtggtcg ggcagagcgg cttgggtaaa
      421 tccaccttaa tcaacaccct cttcaaatcc aaaatcagcc ggaagtcggt gcagcccacc
      481 tcagaggagc gcatccccaa gaccatcgag atcaagtcca tcacgcacga tattgaggag
      541 aaaggcgtcc ggatgaagct gacagtgatt gacacaccag ggttcgggga ccacatcaac
      601 aacgagaact gctggcagcc catcatgaag ttcatcaatg accagtacga gaaatacctg
      661 caggaggagg tcaacatcaa ccgcaagaag cgcatcccgg acacccgcgt ccactgctgc
      721 ctctacttca tccccgccac cggccactcc ctcaggcccc tggacatcga gtttatgaaa
      781 cgcctgagca aggtggtcaa catcgtccct gtcatcgcca aggcggacac actcaccctg
      841 gaggagaggg tccacttcaa acagcggatc accgcagacc tgctgtccaa cggcatcgac
      901 gtgtaccccc agaaggaatt tgatgaggac tcggaggacc ggctggtgaa cgagaagttc
      961 cgggagatga tcccatttgc tgtggtgggc agtgaccacg agtaccaggt caacggcaag
     1021 aggatccttg ggaggaagac caagtggggt accatcgaag ttgaaaacac cacacactgt
     1081 gagtttgcct acctgcggga ccttctcatc aggacgcaca tgcagaacat caaggacatc
     1141 accagcagca tccacttcga ggcgtaccgt gtgaagcgcc tcaacgaggg cagcagcgcc
     1201 atggccaacg gcgtggagga gaaggagcca gaagccccag agatggaaaa cctttacttc
     1261 cagggcagcg gcagcggctg gagccacccg cagttcgaaa aataaactag taataatttt
     1321 gtttaacttt aagaaggaga tatacatatg tcggtcagtg cgagatccgc tgctgctgag
     1381 gagaggagcg tcaacagcag caccatggta gctcaacaga agaaccttga aggctatgtg
     1441 ggatttgcca atctcccaaa tcaagtatac agaaaatcgg tgaagagagg ttttgaattc
     1501 acgcttatgg tagtgggtga atctggattg ggaaagtcga cattaatcaa ctcattattc
     1561 ctcacagatt tgtattctcc agagtatcca ggtccttctc atagaattaa aaagactgta
     1621 caggtggaac aatccaaagt tttaatcaaa gaaggtggtg ttcagttgct gctcacaata
     1681 gttgataccc caggatttgg agatgcagtg gataatagta attgctggca gcctgttatc
     1741 gactacattg atagtaaatt tgaggactac ctaaatgcag aatcacgagt gaacagacgt
     1801 cagatgcctg ataacagggt gcagtgttgt ttatacttca ttgctccttc aggacatgga
     1861 cttaaaccat tggatattga gtttatgaag cgtttgcatg aaaaagtgaa tatcatccca
     1921 cttattgcca aagcagacac actcacacca gaggaatgcc aacagtttaa aaaacagata
     1981 atgaaagaaa tccaagaaca taaaattaaa atatacgaat ttccagaaac agatgatgaa
     2041 gaagaaaata aacttgttaa aaagataaag gaccgtttac ctcttgctgt ggtaggtagt
     2101 aatactatca ttgaagttaa tggcaaaagg gtcagaggaa ggcagtatcc ttggggtgtt
     2161 gctgaagttg aaaatggtga acattgtgat tttacaatcc taagaaatat gttgataaga
     2221 acacacatgc aggacttgaa agatgttact aataatgtcc actatgagaa ctacagaagc
     2281 agaaaacttg cagctgtgac ttataatgga gttgataaca acaagaataa agggcagctg
     2341 actaagagcc ctctggcaca aatggaagaa gaaagaaggg agcatgtagc taaaatgaag
     2401 aagatggaga tggagatgga gcaggtgttt gagatgaagg tcaaagaaaa agttcaaaaa
     2461 ctgaaggact ctgaagctga gctccagcgg cgccatgagc aaatgaaaaa gaatttggaa
     2521 gcacagcaca aagaattgga ggaaaaacgt cgtcagttcg aggatgagaa agcaaactgg
     2581 gaagctcaac aacgtatttt agaacaacag aactcttcaa gaaccttgga aaagaacaag
     2641 aagaaaggga agatctttta aggatcctaa tagtctagag ctagccctag gctgctgcca
     2701 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2761 tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     2821 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg
     2881 ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg aattgttaga cattatttgc
     2941 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
     3001 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     3061 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     3121 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     3181 cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     3241 gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc
     3301 tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga
     3361 agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg
     3421 gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa
     3481 tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg
     3541 ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca
     3601 ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc
     3661 gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc
     3721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3781 gatacatatt tgaatgtatt tagaaaaata aacaaatagc taggtcactc ggtcgctacg
     3841 ctccgggcgt gagactgcgg cgggcgctgc ggacacatac aaagttaccc acagattccg
     3901 tggataagca ggggactaac atgtgaggca aaacagcagg gccgcgccgg tggcgttttt
     3961 ccataggctc cgccctcctg ccagagttca cataaacaga cgcttttccg gtgcatctgt
     4021 gggagccgtg aggctcaacc atgaatctga cagtacgggc gaaacccgac aggacttaaa
     4081 gatccccacc gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg accctgccgt
     4141 ttaccggata cctgttccgc ctttctccct tacgggaagt gtggcgcttt ctcatagctc
     4201 acacactggt atctcggctc ggtgtaggtc gttcgctcca agctgggctg taagcaagaa
     4261 ctccccgttc agcccgactg ctgcgcctta tccggtaact gttcacttga gtccaacccg
     4321 gaaaagcacg gtaaaacgcc actggcagca gccattggta actgggagtt cgcagaggat
     4381 ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc tacactggaa
     4441 ggacagattt ggttgctgtg ctctgcgaaa gccagttacc acggttaagc agttccccaa
     4501 ctgacttaac cttcgatcaa accacctccc caggtggttt tttcgaaggt taagtcagtt
     4561 ggggaactgc ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc
     4621 aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac
     4681 cgctcttgat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     4741 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     4801 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     4861 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4921 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     4981 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     5041 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     5101 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     5161 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     5221 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5281 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5341 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5401 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5461 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5521 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     5581 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     5641 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     5701 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     5761 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     5821 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     5881 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     5941 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     6001 ccataccgcg aaaggttt
//
