LOCUS       sequence-129280-        5782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Sept2 with a YFP fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5782)
  AUTHORS   Spiliotis ET, Kinoshita M, Nelson WJ
  TITLE     A mitotic septin scaffold required for Mammalian chromosome 
            congression and segregation.
  JOURNAL   Science. 2005 Mar 18;307(5716):1781-5.
  PUBMED    15774761
REFERENCE   2  (bases 1 to 5782)
  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: "Science."; 
            date: "2005-03-18"; volume: "307(5716)"; pages: "1781-5"
COMMENT     Sequence Label: pEYFP-N1_SEPT2
FEATURES             Location/Qualifiers
     source          1..5782
                     /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"
     regulatory      1722..1731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1728..1730,1731..1733,1734..2447)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1773..1794)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2034..2053)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2381..2402
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2570..2691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2607..2626)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2661..2680
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2698..3153)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2835..2856)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3047..3066
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3180..3284
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3286..3643
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3286..3477
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3309..3329)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3494..3629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3678..4472
                     /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             3678..4472
                     /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(3732..3751)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4342..4361
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4660..4679)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4704..4751
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5080..5668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5569..5588
                     /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 gctagcgcta
      601 ccggactcag atctcgagcc accatgtcta agcaacaacc aactcagttt ataaatccag
      661 aaactcctgg ctatgttgga tttgcaaatc ttcccaatca agttcaccga aaatcagtga
      721 agaaggggtt cgagttcact ctgatggtgg ttggtgaatc tggtctagga aaatcaactc
      781 tcataaacag cttattcctg actgatctct acccagaaag aattattcct ggagctgcag
      841 agaaaattga aagaactgtc cagatagagg cttcgactgt tgagattgaa gagcggggtg
      901 tgaagctgcg gcttacagta gtggacactc ccggctacgg ggatgccatc aactgcaggg
      961 attgtttcaa gacaattatc tcctacattg atgagcagtt tgaacgctac ctacatgatg
     1021 agagtggact gaacaggcgt cacatcattg ataacagggt acattgttgc ttctacttca
     1081 tttcaccttt tggacatgga ctgaagccct tagatgttgc atttatgaaa gcgatacaca
     1141 ataaggtgaa tattgtgcct gtcattgcga aagctgacac tctcactctg aaggagcgtg
     1201 agcggcttaa gaaaaggatt ttggatgaaa ttgaagagca tagcattaaa atctatcact
     1261 tacctgatgc agagtcagat gaagatgaag actttaagga gcagactaga ctcctcaagg
     1321 ccagtatccc attctctgtg gttggctcca accagttgat tgaagccaaa ggcaagaagg
     1381 ttagaggccg tctctaccca tggggtgttg tagaggtgga gaacccagaa cacaatgact
     1441 ttctgaagct gagaacgatg ctcatcaccc acatgcagga cctacaggaa gtgacccaag
     1501 accttcacta tgaaaacttc cgttctgaga ggctgaagag aggcggcagg aaagtagaga
     1561 atgaggacat gaataaagac cagatcttgc ttgaaaagga ggctgagctc cgccgcatgc
     1621 aagagatgat tgcaagaatg caagcgcaga tgcagatgca gatgcagggt ggtgacagtg
     1681 acagcggggc tctcgggcag catgtgcggg atccaccggt cgccaccatg gtgagcaagg
     1741 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1801 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1861 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1921 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1981 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2041 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2101 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2161 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2221 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2281 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2341 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2401 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     2461 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2521 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2581 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2641 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     2701 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2761 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2821 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2881 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2941 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3001 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3061 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3121 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3181 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3241 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3301 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3481 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3601 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     3661 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3721 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3781 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3841 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3901 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3961 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4021 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4081 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4141 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4201 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4261 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4321 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4381 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4441 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4501 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4561 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4621 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     4681 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     4741 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4801 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4861 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     4921 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5101 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5161 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5221 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5281 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5341 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5401 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5461 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5521 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5581 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5641 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5701 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5761 taaccgtatt accgccatgc at
//
