LOCUS       sequence-349914-        6066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6066)
  AUTHORS   Gilden JK, Peck S, Chen YC, Krummel MF
  TITLE     The septin cytoskeleton facilitates membrane retraction during 
            motility and blebbing.
  JOURNAL   J Cell Biol. 2012 Jan 9;196(1):103-14.
  PUBMED    22232702
REFERENCE   2  (bases 1 to 6066)
  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 
            Biol."; date: "2012-01-9"; volume: "196(1)"; pages: "103-14"
COMMENT     Sequence Label: pMK782 Septin6-GFP
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..292
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     249..269
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(294..398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      425..880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(512..531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     722..743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    887..1008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(898..917)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     952..971
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2533..3243,3244..3246,3247..3249))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2533..3243,3244..3246,3247..3249))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2575..2596)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2924..2943
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3183..3204
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3246..3255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3294..3497)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3323..3343)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3498..3801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3976..4564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4056..4075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4893..4940
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4965..4984
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5172..5966)
                     /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             complement(5172..5966)
                     /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(5283..5302)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5893..5912
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6001..6066,1..292))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6015..6066,1..84)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
       61 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      121 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      241 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
      301 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      421 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
      481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
      541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
      601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
      661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
      721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
      781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
      841 tttaacgcga attttaacaa aatattaacg cttacaattt acgcgttaag atacattgat
      901 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
      961 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1021 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1081 aacctctaca aatgtggtat ggctgattat gatcagttat ctagatccgg tggatcctgc
     1141 ggccgcttaa aaaaagttct tcttctcttt gtcccttttc agagtctgtg agcccccagc
     1201 ctgagagccc tgagactgga gcagctcagc tgccgccttt ctctgcttga aggcgttcat
     1261 ttcctcatcc agacatttct tcttatcctc cagcttcttc ttctcttcct ggtgcagttt
     1321 cttcagacgg tcaaatttct catgcagctc tttttcagcc tccttgagct ccgcttcttt
     1381 ctctttgact ctctggacaa acatctgcct catctcctct tctttcttct ggagctcccc
     1441 aaggaactca tttctctttg cctcatatgt ctcctgcaaa ctgaatggtt tgctgtcagg
     1501 gtcggtgtcc ttgaagccca tctcctccag cttacagcgg cggtacaatt catagtgccg
     1561 ggcgtgggtc tgctcccgca gatcctccat gttgacccga atcagcatct cccgcagctt
     1621 cacaaagtcg cagtgagcct cgttttcaac ctgcacagtc ccccaaggat actgccgagc
     1681 ccgcatcatc ttgttgccta tcttcacttc ttctgtgctg ccaacaacag caaacggcag
     1741 gtgggcattc atggttccat tgatctcgct caccgactca tcgtctgtgg ggaactgata
     1801 gatctggact ccattgctca caagttcgct ggtgattttg atcttgaact ttgccagctc
     1861 actcttagaa atggcatctg acttggcaat aacgggaatg atgttcacct tactatccag
     1921 cttcttcatg gtcactaggt ccagagactt aagcgaatgt cctgtcgggg caatgaaata
     1981 caagcagaca tggattcggg agtcatggta ggagtgcagt actcttcgga tcttcagctc
     2041 ctcttgtagg taggcctcaa actgagcatc aatgaattcc acgatgggct tgtagctgtc
     2101 ctccttgttg atctgatccc caaagcctac agtgctcaca attgtgagtt tcaaccccac
     2161 gttgctctcc tggaggtcgt aggtattgga ctggagctga acccctggct gtgtgtgggt
     2221 cgctggctcc ccttcaaact tggtgttaaa cagggtatcc atgagcgtgg acttgcccaa
     2281 acctgtctcc cccacgcaca ggatgttgaa gcagaagccc tggctgactg acttattcac
     2341 cagctggtca ggcaagctgt cgaaccccac atgtccagcc agggggacag ttcgacaatc
     2401 ttcgcccacc tggcgagcta tatcggccgc tgccatcaga atacctgcgg agcctccccc
     2461 accgcttgcc agagcagacc caccgcctcc actgatagcc agcgaagctt gagctcgaga
     2521 tctgagtccg gacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     2581 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
     2641 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2701 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     2761 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     2821 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2881 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2941 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     3001 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     3061 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     3121 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     3181 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     3241 gctcaccatg gtggcgaccg gtagcgctag cggatctgac ggttcactaa accagctctg
     3301 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     3361 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     3421 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     3481 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     3541 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     3601 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     3661 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     3721 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     3781 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     3841 attgattact attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga
     3901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4621 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     4741 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     4801 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     4861 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     4921 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     4981 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     5041 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     5101 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     5161 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     5221 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     5281 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     5341 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     5401 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     5461 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     5521 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     5581 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     5641 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     5701 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     5761 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     5821 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     5881 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     5941 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     6001 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     6061 cgaggc
//
