LOCUS       sequence-331147-        6943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PRD-deleted DNM2 mutant (aa 1-742) in pEGFP-N1 vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6943)
  AUTHORS   Cao F, Zhou Y, Liu X, Yu CH
  TITLE     Podosome formation promotes plasma membrane invagination and 
            integrin-beta3 endocytosis on a viscous RGD-membrane.
  JOURNAL   Commun Biol. 2020 Mar 13;3(1):117. doi: 10.1038/s42003-020-0843-2.
  PUBMED    32170110
REFERENCE   2  (bases 1 to 6943)
  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: "Commun 
            Biol."; date: "2020-03-13"; volume: "3(1):117. doi"; pages: " 
            10.1038/s42003-020-0843-2"
COMMENT     Sequence Label: DNM2 delta PRD
FEATURES             Location/Qualifiers
     source          1..6943
                     /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"
     misc_feature    2823..2875
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2883..2892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2889..2891,2892..2894,2895..3608)
                     /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(2889..2891,2892..2894,2895..3605)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2934..2955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3195..3214)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3542..3563
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3731..3852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3768..3787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3822..3841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3859..4314)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3996..4017)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4208..4227
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4341..4445
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4447..4804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4447..4638
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4470..4490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4655..4790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4839..5633
                     /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             4839..5633
                     /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(4893..4912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5503..5522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5821..5840)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5865..5912
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6241..6829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6730..6749
                     /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 gcaaccgcgg gatggaagag ctgatcccgc tggtcaacaa actgcaggac gccttcagct
      661 ccatcggcca gagctgccac ctggacctgc cgcagatcgc tgtagtgggc ggccagagcg
      721 ccggcaagag ctcggtgctg gagaacttcg tgggccggga cttccttccc cgcggttcag
      781 gaatcgtcac ccggcggcct ctcattctgc agctcatctt ctcaaaaaca gaacatgccg
      841 agtttttgca ctgcaagtcc aaaaagttta cagactttga tgaagtccgg caggagattg
      901 aagcagagac cgacagggtc acggggacca acaaaggcat ctccccagtg cccatcaacc
      961 ttcgagtcta ctcgccacac gtgttgaact tgaccctcat cgacctcccg ggtatcacca
     1021 aggtgcctgt gggcgaccag cctccagaca tcgagtacca gatcaaggac atgatcctgc
     1081 agttcatcag ccgggagagc agcctcattc tggctgtcac gcccgccaac atggacctgg
     1141 ccaactccga cgccctcaag ctggccaagg aagtcgatcc ccaaggccta cggaccatcg
     1201 gtgtcatcac caagcttgac ctgatggacg agggcaccga cgccagggac gtcttggaga
     1261 acaagttgct cccgttgaga agaggctaca ttggcgtggt gaaccgcagc cagaaggata
     1321 ttgagggcaa gaaggacatc cgtgcagcac tggcagctga gaggaagttc ttcctctccc
     1381 acccggccta ccggcacatg gccgaccgca tgggcacgcc acatctgcag aagacgctga
     1441 atcagcaact gaccaaccac atccgggagt cgctgccggc cctacgtagc aaactacaga
     1501 gccagctgct gtccctggag aaggaggtgg aggagtacaa gaactttcgg cccgacgacc
     1561 ccacccgcaa aaccaaagcc ctgctgcaga tggtccagca gtttggggtg gattttgaga
     1621 agaggatcga gggctcagga gatcaggtgg acactctgga gctctccggg ggcgcccgaa
     1681 tcaatcgcat cttccacgag cggttcccat ttgagctggt gaagatggag tttgacgaga
     1741 aggacttacg acgggagatc agctatgcca ttaagaacat ccatggagtc aggaccgggc
     1801 ttttcacccc ggacttggca ttcgaggcca ttgtgaaaaa gcaggtcgtc aagctgaaag
     1861 agccctgtct gaaatgtgtc gacctggtta tccaggagct aatcaataca gttaggcagt
     1921 gtaccagtaa gctcagttcc tacccccggt tgcgagagga gacagagcga atcgtcacca
     1981 cttacatccg ggaacgggag gggagaacga aggaccagat tcttctgctg atcgacattg
     2041 agcagtccta catcaacacg aaccatgagg acttcatcgg gtttgccaat gcccagcaga
     2101 ggagcacgca gctgaacaag aagagagcca tccccaatca ggtgatccgc aggggctggc
     2161 tgaccatcaa caacatcagc ctgatgaaag gcggctccaa ggagtactgg tttgtgctga
     2221 ctgccgagtc actgtcctgg tacaaggatg aggaggagaa agagaagaag tacatgctgc
     2281 ctctggacaa cctcaagatc cgtgatgtgg agaagggctt catgtccaac aagcacgtct
     2341 tcgccatctt caacacggag cagagaaacg tctacaagga cctgcggcag atcgagctgg
     2401 cctgtgactc ccaggaagac gtggacagct ggaaggcctc gttcctccga gctggcgtct
     2461 accccgagaa ggaccaggca gaaaacgagg atggggccca ggagaacacc ttctccatgg
     2521 acccccaact ggagcggcag gtggagacca ttcgcaacct ggtggactca tacgtggcca
     2581 tcatcaacaa gtccatccgc gacctcatgc caaagaccat catgcacctc atgatcaaca
     2641 atacgaaggc cttcatccac cacgagctgc tggcctacct atactcctcg gcagaccaga
     2701 gcagcctcat ggaggagtcg gctgaccagg cacagcggcg ggacgacatg ctgcgcatgt
     2761 accatgccct caaggaggcg ctcaacatca tcggtgacat cagcaccagc actgtgtcca
     2821 cgctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
     2881 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2941 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3001 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3061 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3121 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3181 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3241 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3301 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     3361 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3421 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3481 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3541 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3601 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3661 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3721 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3841 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3901 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3961 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4021 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4081 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4141 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4201 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4261 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4381 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4441 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4501 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4621 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4681 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4741 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4801 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4861 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4921 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4981 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5041 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5101 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5161 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5221 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5281 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5341 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5401 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5461 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5521 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5581 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5641 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5701 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5761 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5821 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5881 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5941 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6001 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6061 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6121 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6181 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6361 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6421 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6901 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
