LOCUS       sequence-389673-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of GFP-chimera proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7319)
  AUTHORS   Jimah JR, Kundu N, Stanton AE, Sochacki KA, Canagarajah B, Chan L, 
            Strub MP, Wang H, Taraska JW, Hinshaw JE
  TITLE     Cryo-EM structures of membrane-bound dynamin in a post-hydrolysis 
            state primed for membrane fission.
  JOURNAL   Dev Cell. 2024 Apr 23:S1534-5807(24)00233-8. doi: 
            10.1016/j.devcel.2024.04.008.
  PUBMED    38663399
REFERENCE   2  (bases 1 to 7319)
  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: "Dev Cell. 
            2024 Apr 23:S1534-5807(24)00233-8. doi: 
            10.1016/j.devcel.2024.04.008."
COMMENT     Sequence Label: Dyn1 K44A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..133)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    158..205
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      534..1122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1023..1042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1297..1600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1601..1804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1755..1775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4495..4504
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4501..4503,4504..4506,4507..5220)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4501..4503,4504..4506,4507..5217)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4546..4567)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4807..4826)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5154..5175
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5343..5464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5380..5399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5434..5453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5471..5926)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5608..5629)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5820..5839
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5953..6057
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6059..6416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6059..6250
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6082..6102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6267..6402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6451..7245
                     /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             6451..7245
                     /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(6505..6524)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7115..7134
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
       61 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
      121 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
      181 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
      241 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
      301 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
      361 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
      421 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      481 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      541 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      601 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      661 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
      721 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      781 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      841 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      901 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      961 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1021 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1081 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1141 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1201 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     1261 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1321 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1381 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1441 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     1501 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     1561 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1621 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1681 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     1741 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     1801 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatcc tgccatgggc
     1861 aaccgcggca tggaagatct catcccgctg gtcaaccggc tgcaagacgc cttctctgcc
     1921 atcggccaga acgcggacct cgacctgccg cagatcgctg tggtgggcgg ccagagcgcc
     1981 ggcgcgagct cggtgctcga gaatttcgta ggcagggact tcttgcctcg aggatctggc
     2041 attgttaccc gacgtcccct ggtcttgcag ctggtcaatg caaccacaga atatgccgag
     2101 ttcctgcact gcaagggaaa gaaattcacc gacttcgagg aggtgcgcct tgagatcgag
     2161 gccgagaccg acagggtcac cggcaccaac aagggcatct cgccggtgcc tatcaacctc
     2221 cgcgtctact cgccgcacgt gctgaacctg accctggtgg acctgcccgg aatgaccaag
     2281 gtcccggtgg gggaccaacc tcccgacatc gagttccaga tccgagacat gcttatgcag
     2341 tttgtcacca aggagaactg cctcatcctg gccgtgtccc ccgccaactc tgacctggcc
     2401 aattctgacg ccctcaaggt cgccaaggag gtggaccccc agggccagcg caccatcggg
     2461 gtcatcacca agctggacct gatggacgag ggcacagatg cccgtgatgt gctggagaac
     2521 aagctgctcc ccctgcgcag aggctacatt ggagtggtga accggagcca gaaggacatt
     2581 gatggcaaga aggacattac cgccgccttg gctgctgaac gaaagttctt cctctcccat
     2641 ccatcttatc gccacttggc tgaccgtatg ggcacgccct acctgcagaa ggtcctcaat
     2701 cagcaactga cgaaccacat ccgggacaca ctgccggggc tgcggaacaa gctgcagagc
     2761 cagctactgt ccattgagaa ggaggtggag gaatacaaga acttccgccc tgatgaccca
     2821 gctcgcaaga ccaaggccct gctgcagatg gtccagcagt ttgccgtaga ctttgagaag
     2881 cgcattgagg gctcaggaga tcagatcgac acctacgaac tgtcaggggg agcccgcatt
     2941 aaccgaatct tccacgagcg cttccctttc gagctggtca agatggagtt tgatgagaag
     3001 gaactccgaa gggagatcag ctatgctatc aagaatatcc atggcattag aacggggctg
     3061 tttaccccag acatggcctt tgagaccatt gtgaaaaagc aggtgaagaa gatccgagaa
     3121 ccgtgtctca agtgtgtgga catggttatc tcggagctaa tcagcaccgt tagacagtgc
     3181 accaagaagc tccagcagta cccgcggcta cgggaggaga tggagcgcat cgtgaccacc
     3241 cacatccggg agcgcgaggg ccgcactaag gagcaggtca tgcttctcat cgatatcgag
     3301 ctggcttaca tgaacaccaa ccatgaggac ttcataggct ttgccaatgc tcagcagagg
     3361 agcaaccaga tgaacaagaa gaagacttca gggaaccagg atgagattct ggtcatccgc
     3421 aagggctggc tgactatcaa taatattggc atcatgaaag ggggctccaa ggagtactgg
     3481 tttgtgctga ctgctgagaa tctgtcctgg tacaaggatg atgaggagaa agagaagaaa
     3541 tacatgctgt ctgtggacaa cctcaagctg cgggacgtgg agaagggctt tatgtcgagc
     3601 aagcatatct ttgccctctt taacacggag cagaggaatg tctacaagga ttatcggcag
     3661 ctggagctag cctgtgagac acaggaggag gtggacagct ggaaggcctc cttcctgagg
     3721 gctggcgtgt accctgagcg tgttggggac aaagagaaag ccagcgagac cgaggagaat
     3781 ggctccgaca gcttcatgca ttccatggac ccacagctgg aacggcaagt ggagaccatc
     3841 cggaatcttg tggactcata catggccatt gtcaacaaga ccgtgaggga cctcatgccc
     3901 aagaccatca tgcacctcat gattaacaat accaaggagt tcatcttctc ggagctgctg
     3961 gccaacctgt actcgtgtgg ggaccagaac acgctgatgg aggagtcggc ggagcaggca
     4021 cagcggcgcg acgagatgct gcgcatgtac cacgcactga aggaggcgct cagcatcatc
     4081 ggcaacatca acacgaccac cgtcagcacg cccatgcccc cgcccgtgga cgactcctgg
     4141 ctgcaggtgc agagcgtacc ggccggacgc aggtcgccca cgtccagccc cacgccgcag
     4201 cgccgagccc ccgccgtgcc cccagcccgg cccgggtcgc ggggccctgc tcctgggcct
     4261 ccgcctgctg ggtccgccct ggggggggcg ccccccgtgc cctccaggcc gggggcttcc
     4321 cctgaccctt tcggccctcc ccctcaggtg ccctcgcgcc ccaaccgcgc cccgcccggg
     4381 gtccccagcc gatcgggtca ggcaagtcca tcccgtcctg agagccccag gccccccttc
     4441 gacaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcgccacc
     4501 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4561 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4621 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4681 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     4741 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4801 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4861 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4921 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     4981 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     5041 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5101 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5161 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     5221 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     5281 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     5341 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5401 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5461 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5521 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5581 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5641 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5701 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5761 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     5821 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     5881 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     5941 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6001 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     6061 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     6121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     6181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     6301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     6421 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     6481 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6541 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6601 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     6661 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6721 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6781 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6841 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6901 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6961 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     7021 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     7081 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     7141 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     7201 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     7261 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgcc
//
