LOCUS       sequence-358038-        6410 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mammalian expression of human SEPT9_i3 fused to monomeric Apple and 
            CAAX (HRas).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6410)
  AUTHORS   Martins CS, Taveneau C, Castro-Linares G, Baibakov M, Buzhinsky N, 
            Eroles M, Milanovic V, Omi S, Pedelacq JD, Iv F, Bouillard L, 
            Llewellyn A, Gomes M, Belhabib M, Kuzmic M, Verdier-Pinard P, Lee S,
            Badache A, Kumar S, Chandre C, Brasselet S, Rico F, Rossier O, 
            Koenderink GH, Wenger J, Cabantous S, Mavrakis M
  TITLE     Human septins organize as octamer-based filaments and mediate 
            actin-membrane anchoring in cells.
  JOURNAL   J Cell Biol. 2023 Mar 6;222(3):e202203016. doi: 
            10.1083/jcb.202203016. Epub 2022 Dec 23.
  PUBMED    36562751
REFERENCE   2  (bases 1 to 6410)
  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 
            Biol."; date: "2023-03-6"; volume: "222(3):e202203016. doi"; pages: 
            " 10.1083/jcb.202203016. Epub 2022 Dec 23"
COMMENT     Sequence Label: pCMV_SEPT9_i3-mApple-CAAX
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2298..3005
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2711..2730
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3198..3319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3235..3254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3289..3308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3326..3781)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3463..3484)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3675..3694
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3808..3912
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3914..4271
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3914..4105
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3937..3957)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4122..4257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4306..5100
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4306..5100
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4360..4379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4970..4989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5288..5307)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5332..5379
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5708..6296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6197..6216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 agagggaccg gatctcagcc ttgaaaagat cttttgaggt cgaggaggtc gagacaccca
      661 actccacccc accccggagg gtccagactc ccctactccg agccactgtg gccagctcca
      721 cccagaaatt ccaggacctg ggcgtgaaga actcagaacc ctcggcccgc catgtggact
      781 ccctaagcca acgctccccc aaggcgtccc tgcggagggt ggagctctcg ggccccaagg
      841 cggccgagcc ggtgtcccgg cgcactgagc tgtccattga catctcgtcc aagcaggtgg
      901 agaacgccgg ggccatcggc ccgtcccggt tcgggctcaa gagggccgag gtgttgggcc
      961 acaagacgcc agaaccggcc cctcggagga cggagatcac catcgtcaaa ccccaggagt
     1021 cagcccaccg gaggatggag ccccctgcct ccaaggtccc cgaggtgccc actgcccctg
     1081 ccaccgacgc agcccccaag agggtggaga tccagatgcc caagcctgct gaggcgccca
     1141 ccgcccccag cccagcccag accttggaaa attcagagcc tgcccctgtg tctcagctgc
     1201 agagcaggct ggagcccaag ccccagcccc ctgtggctga ggctacaccc cggagccagg
     1261 aggccactga ggcggctccc agctgcgttg gcgacatggc cgacaccccc agagatgccg
     1321 ggctcaagca ggcgcctgca tcacggaacg agaaggcccc ggtggacttc ggctacgtgg
     1381 ggattgactc catcctggag cagatgcgcc ggaaggccat gaagcagggc ttcgagttca
     1441 acatcatggt ggtcgggcag agcggcttgg gtaaatccac cttaatcaac accctcttca
     1501 aatccaaaat cagccggaag tcggtgcagc ccacctcaga ggagcgcatc cccaagacca
     1561 tcgagatcaa gtccatcacg cacgatattg aggagaaagg cgtccggatg aagctgacag
     1621 tgattgacac accagggttc ggggaccaca tcaacaacga gaactgctgg cagcccatca
     1681 tgaagttcat caatgaccag tacgagaaat acctgcagga ggaggtcaac atcaaccgca
     1741 agaagcgcat cccggacacc cgcgtccact gctgcctcta cttcatcccc gccaccggcc
     1801 actccctcag gcccctggac atcgagttta tgaaacgcct gagcaaggtg gtcaacatcg
     1861 tccctgtcat cgccaaggcg gacacactca ccctggagga gagggtccac ttcaaacagc
     1921 ggatcaccgc agacctgctg tccaacggca tcgacgtgta cccccagaag gaatttgatg
     1981 aggactcgga ggaccggctg gtgaacgaga agttccggga gatgatccca tttgctgtgg
     2041 tgggcagtga ccacgagtac caggtcaacg gcaagaggat ccttgggagg aagaccaagt
     2101 ggggtaccat cgaagttgaa aacaccacac actgtgagtt tgcctacctg cgggaccttc
     2161 tcatcaggac gcacatgcag aacatcaagg acatcaccag cagcatccac ttcgaggcgt
     2221 accgtgtgaa gcgcctcaac gagggcagca gcgccatggc caacggcgtg gaggagaagg
     2281 agccagaagc cccagagatg gtgagcaagg gcgaggagaa taacatggcc atcatcaagg
     2341 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     2401 agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca
     2461 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2521 aggtctacat taagcaccca gccgacatcc ccgactactt caagctgtcc ttccccgagg
     2581 gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg cattattcac gttaaccagg
     2641 actcctccct gcaggacggc gtgttcatct acaaggtgaa gctgcgcggc accaacttcc
     2701 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc gaggagcgga
     2761 tgtaccccga ggacggcgcc ctgaagagcg agatcaagaa gaggctgaag ctgaaggacg
     2821 gcggccacta cgccgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2881 ccggcgccta catcgtcgac atcaagttgg acatcgtgtc ccacaacgag gactacacca
     2941 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gatgagctgt
     3001 acaagaagct gaaccctcct gatgagagtg gccccggctg catgagctgc aagtgtgtgc
     3061 tctcctaagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3121 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3181 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3301 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3361 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3421 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3481 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3541 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3601 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3661 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3721 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3781 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3841 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3901 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4201 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4261 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4321 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4381 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4441 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4501 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4561 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4621 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4681 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4741 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4801 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4861 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4921 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4981 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5041 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5101 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5161 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5221 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5281 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5341 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5401 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5461 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5521 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5581 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5641 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5701 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5761 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5821 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5881 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5941 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6001 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6061 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6121 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6181 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6241 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6301 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6361 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
