LOCUS       sequence-250220-        7331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Dynamin 2 K44A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7331)
  AUTHORS   Chua J, Rikhy R, Lippincott-Schwartz J
  TITLE     Dynamin 2 orchestrates the global actomyosin cytoskeleton for 
            epithelial maintenance and apical constriction.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Dec 8;106(49):20770-5. doi: 
            10.1073/pnas.0909812106. Epub 2009 Nov 30.
  PUBMED    19948954
REFERENCE   2  (bases 1 to 7331)
  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"; doi: 
            "10.1073/pnas.0909812106"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2009-12-8- 8"; volume: "106"; issue: "49"; pages: "20770-5"
COMMENT     Sequence Label: RFP Dynamin2 K44A
FEATURES             Location/Qualifiers
     source          1..7331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1297..1308
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3253..3262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3259..3936
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3259..3936
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3389..3407)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3657..3676
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3851..3874
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4059..4180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4096..4115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4150..4169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4187..4642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4324..4345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4536..4555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4669..4773
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4775..5132
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4775..4966
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4798..4818)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4983..5118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5167..5961
                     /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             5167..5961
                     /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(5221..5240)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5831..5850
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6149..6168)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6193..6240
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6569..7157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7058..7077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttggc accatgggca accgcgggat ggaagagctc
      601 atcccgctgg ttaacaagct gcaggacgcc ttcagttcca tcggtcagag ctgccacctg
      661 gacctgccgc agatcgccgt ggtgggcggc cagagcgccg gcgcgagttc ggtgctcgag
      721 aacttcgtgg gccgggactt ccttccacga ggatcaggaa ttgtcacccg gaggcctctc
      781 attctgcagc tcatcttctc caaaacagaa tatgcggagt ttttgcactg caagtccaaa
      841 aagtttacag actttgatga agtccggcag gagatcgaag cagagactga ccgggtcaca
      901 ggcaccaaca aaggcatctc cccggtgccc atcaaccttc gggtctattc accacacgtg
      961 ttgaacttga ccctcatcga cctcccaggc atcactaagg tgccagtggg ggaccagcca
     1021 ccagacatcg agtaccagat caaggacatg atcctgcagt ttatcagccg ggagagcagc
     1081 ctcattcttg ccgtcacacc tgccaacatg gacttggcca actcggatgc cctcaagctg
     1141 gccaaggagg tggaccccca aggcctgcgg accattggtg tcatcaccaa gctagacctg
     1201 atggatgaag gcacagatgc cagggatgtc ctggagaaca agctcctgcc cttgagaaga
     1261 ggctacatcg gcgtggttaa ccgcagccag aaagacatcg agggcagaaa ggacatccgg
     1321 gcagctctgg cagctgagag gaaattcttc ctctcccacc cagcctaccg gcacatggct
     1381 gaccgcatgg gtaccccaca cttgcagaaa acactgaacc agcaactgac caaccacatc
     1441 cgggagtcac tgccaaccct gcgcagcaag ctgcagagcc aactgctgtc cctggagaag
     1501 gaagtggaag agtacaagaa tttccggcct gacgacccca cacgcaagac caaagccctg
     1561 ctgcagatgg tccagcagtt tggagtggac tttgagaagc gaatcgaagg ctctggagat
     1621 caagtcgaca ccctggagct gtctgggggc gcccgcatca atcgcatctt ccacgagcgc
     1681 tttccctttg agctggtgaa gatggagttt gacgagaaag acctccgaag agagatcagc
     1741 tacgccatta agaacatcca cggagtcagg acggggctct tcacccccgg catggccttt
     1801 gaagccatcg tgaaaaaaca acttgtaaag ctcaaagagc cgagtttgaa gtgtgttgac
     1861 ctggtggtgt cggagctagc cacggtcatt aaaaagtgtg ctgagaagct cagttcctac
     1921 ccccgccttc gagaggagac cgaacgcatt gtcaccacct acatcaggga gcgagaaggg
     1981 agaaccaagg accagatcct tcttctgatt gacattgagc agtcctacat caacacaaac
     2041 catgaagact tcattggatt tgccaatgcc cagcagagga gcacgcagct gaacaagaag
     2101 agggccatac ctaaccaggg ggagatcttg gtgatccgca ggggctggtt gaccatcaac
     2161 aacatcagct tgatgaaagg cggttccaag gagtactggt tcgtgctgac tgctgagtcg
     2221 ttgtcttggt acaaggatga agaggaaaaa gaaaagaagt acatgctgcc actagacaac
     2281 ctcaaaatac gggatgtgga gaagggcttc atgtccaaca agcatgtgtt tgccatcttc
     2341 aacacagagc agaggaacgt ctacaaggac ctgcgacaga tcgaactggc ttgtgactcc
     2401 caggaagatg tggacagctg gaaggcttcg ttcctgcggg ctggggtcta cccagagaag
     2461 gaccaggcag agaacgagga tggagcacaa gagaacacct tctccatgga cccgcagctg
     2521 gagcggcagg tggagaccat ccgcaacttg gtggactcct atgcggccat catcaacaag
     2581 tccatccggg acctcatgcc aaagaccatc atgcacctca tgatcaacaa cacaaaggcc
     2641 ttcatccgcc atgagctgct agcctacctg tactcatcag cagaccagag cagcctcatg
     2701 gaggagtcag ctgagcaggc tcagcggcgg gacgatatgc tgcgcatgta ccatgcactc
     2761 aaggaagcgc tcaacatcat tggggacatc agcaccagca ccgtgtccac gcctgtgccc
     2821 ccacctgtcg atgatacttg gctccggaac accagcagcc acagccccac tccacagcgc
     2881 agacctgtgt ccagtgtgca cccgccgggc cgcccacccg cagtgcgggg ccccacaccg
     2941 gggcctcccc tcattcctat gcctgtgggg gccacatcct ccttctcggc accccccatc
     3001 ccatcccggc ctggaccaca gaatgtgttt gccaacaatg accccttctc tgcgccacct
     3061 cagataccgt ctcggccagc acggattccc cctgggatcc ctccaggagt gcccagcaga
     3121 agagcgcctg ctgcgcccag ccggcccacc attatccgcc cagccgagcc gtccctgctc
     3181 gacgctgcag gggacagtgt tctgggggga attctgcagt cgacggtacc gcgggcccgg
     3241 gatccaccgg tcgccaccat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc
     3301 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     3361 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     3421 cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc ctacgtgaag
     3481 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     3541 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     3601 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     3661 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta ccccgaggac
     3721 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac
     3781 gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag
     3841 accgacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3901 gagcgcgccg agggccgcca ctccaccggc gcctaaagcg gccgcgactc tagatcataa
     3961 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4021 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4081 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4141 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     4201 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4261 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4321 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4381 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4441 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4501 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4561 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4621 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4741 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4801 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4861 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4921 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4981 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5041 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5101 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5161 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5221 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5281 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5341 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5401 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5461 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5521 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5581 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5641 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5701 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5761 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5821 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5881 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5941 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6001 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6061 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6121 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6181 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6241 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6301 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6361 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6421 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6481 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6541 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6601 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6661 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6721 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6781 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6841 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6901 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6961 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7021 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7081 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7141 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7201 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7261 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     7321 atggagttcc g
//
