LOCUS       sequence-203441-        8449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donor vector for knock-in gene editing at human DNM1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8449)
  AUTHORS   Srinivasan S, Burckhardt CJ, Bhave M, Chen Z, Chen PH, Wang X, 
            Danuser G, Schmid SL
  TITLE     A noncanonical role for dynamin-1 in regulating early stages of 
            clathrin-mediated endocytosis in non-neuronal cells.
  JOURNAL   PLoS Biol. 2018 Apr 18;16(4):e2005377. doi: 
            10.1371/journal.pbio.2005377. eCollection 2018 Apr.
  PUBMED    29668686
REFERENCE   2  (bases 1 to 8449)
  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: "PLoS 
            Biol."; date: "2018-04-18"; volume: "16(4):e2005377. doi"; pages: " 
            10.1371/journal.pbio.2005377. eCollection 2018 Apr"
COMMENT     Sequence Label: pRS DV DNM1-EGFP
FEATURES             Location/Qualifiers
     source          1..8449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1378..1399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1590..1609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1826..1848
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1840..1857
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1867..1886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2928..2930,2931..3644)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2970..2991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3231..3250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3578..3599
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3767..3888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3804..3823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3858..3877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4847..4867)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4847..4865)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4886..4902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4886..4902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4899..4921)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4910..4926
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4934..4940,4941..4958,4959..4964))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5117..5134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5288..5876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5368..5387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6047..6838,6839..6907))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6670..6689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6908..7012)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7039..8381
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(8002..8049)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     8422..8440
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtacca cgcagtgtag gagccgcgtc
     1921 tgaaaagcca caccagctca cagtcccagc agggcccggt ggggcggcca gggcggcaca
     1981 ggcatcaggt ccccacctcc ttccctcttt gcctactcgc agaccaagga gttcatcttc
     2041 tcggagctgc tggccaacct gtactcgtgt ggggaccaga acacgctgat ggaggagtcg
     2101 gcggagcagg cacagcggcg cgacgagatg ctgcgcatgt accacgcact gaaggaggcg
     2161 ctcagcatca tcggcgacat caacacgacc accgtcagca cgcccatgcc cccgcccgtg
     2221 gacgactcct ggctgcaggt gcagagcgta ccggccggac gcaggtacca gggccggccc
     2281 ccacggcccc aaagcccccc agcccggggc ccgcgggagg aatgccggga ccgggcagtg
     2341 gcgcgcccgc gtcaccgggg tggctcccac ctggagcgag gggcggagct tagagagggc
     2401 ggggcttgtc gcggggcggg gcttgccgtg gagagctggc tgcaaggctg ggtggagctg
     2461 gggcgttggc gccgctgggg gcggggctta agctccggcg accgccttag gggtgcggca
     2521 gggaggggct tgcgtgcatg ggcgtggcca gcactgggct ggggcggggc ctctgggtgg
     2581 gcggagctgc tcatctcgcc tctccttgtt cctcgctccc tgtcgccctc aggtcgccca
     2641 cgtccagccc cacgccgcag cgccgagccc ccgccgtgcc cccagcccgg cccgggtcgc
     2701 ggggccctgc tcctgggcct ccgcctgctg ggtccgccct ggggggggcg ccccccgtgc
     2761 cctccaggcc gggggcttcc cctgaccctt tcggccctcc ccctcaggtg ccctcgcgcc
     2821 ccaaccgcgc cccgcccggg gtccccagcc gatcgggtca ggcaagtcca tcccgtcctg
     2881 agagccccag gccccccttc gacctcgatc ccccagtcgc gaccttagtg agcaagggcg
     2941 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3001 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3061 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     3121 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3181 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3241 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3301 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3361 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3421 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3481 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3541 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3601 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     3661 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3721 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3781 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3841 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttagc tgaatgcggc
     3901 tggagaggct gccggacggg cgtggccggg agggaaatgg ggctggattc cagagcatcg
     3961 gacctggccg ccagaaccgg ccgtttctat cccaggcaat cggactctgg gtgccggagc
     4021 cacgccacgt gtggccgagg gctggcggag cctgcccccg agggagcgct gagtcccgga
     4081 ggtggtcgtt tctggggagg gggctgtggg gctcgtccca cctgccccct tctttccagc
     4141 acttgcatgg cgcttccctc tattttcact cttggcggcc gcccacacgt tgcattcctc
     4201 ctcctttctt cttcttgctg tcctccatcc tccattccgt ccaattcctc tcccagcccc
     4261 tggggagcct accttaggtc tgactctgaa ccccgatctg ctctgagtgt gtggatttcc
     4321 ttcagctacc ctgatgtccc cacttcccag ccctgactcc tttgagccat cccagggggt
     4381 gtccagccac tggcccacag gagcagaggc caggctgtga ctgtgtgact agaaaggtgt
     4441 gtgatgtgtg tgtgggcgtg cacacgagtg tgagagtgta agcatggcac ccaggcccgg
     4501 gctaggacag gagcagctgg ggaagcagcc tgggccgtgg gcaaactgat ggcttttttc
     4561 ctccccccaa ccaccccctc cgcgacttct ggccctaccc agtctaaacc aatgtggtag
     4621 gggtggcggc aggggcaggg atgggagtgc ttgaagcctg cttcattccc aaagatttct
     4681 agggaaaagc tttctactac atcctttggc ttgaccttct tgacccatga ctctctttca
     4741 aggaagttca ctccaattcc cagtctgccc tgtttccact ggccacattc tctaaggaag
     4801 aacacagcag gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     4861 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4921 tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4981 gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5041 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5101 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5161 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5221 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5281 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5341 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5401 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5461 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5521 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5581 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5641 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5701 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5761 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5821 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5881 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5941 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6001 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6061 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6181 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7021 cccgaaaagt gccacctgaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     7081 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     7141 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     7201 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     7261 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     7321 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     7381 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     7441 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     7501 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     7561 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     7621 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     7681 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     7741 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     7801 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     7861 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     7921 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct
     7981 tttggttttc aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat
     8041 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac
     8101 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat
     8161 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg
     8221 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt
     8281 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta
     8341 gtctcatcct tcaatgctat catttccttt gatattggat catattaaga aaccattatt
     8401 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
