LOCUS       sequence-314040-        7319 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 7319)
  AUTHORS   Song BD, Yarar D, Schmid SL
  TITLE     An assembly-incompetent mutant establishes a requirement for dynamin
            self-assembly in clathrin-mediated endocytosis in vivo.
  JOURNAL   Mol Biol Cell. 2004 May;15(5):2243-52. Epub 2004 Mar 5.
  PUBMED    15004222
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: "Mol Biol 
            Cell."; date: "2004-05"; volume: "15(5)"; pages: "2243-52. Epub 2004
            Mar 5"
COMMENT     Sequence Label: K44A dynamin 1 pEGFP
FEATURES             Location/Qualifiers
     source          1..7319
                     /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"
     regulatory      3199..3208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3205..3207,3208..3210,3211..3924)
                     /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(3205..3207,3208..3210,3211..3921)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3250..3271)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3511..3530)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3858..3879
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4047..4168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4084..4103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4138..4157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4175..4630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4312..4333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4524..4543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4763..5120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4763..4954
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4786..4806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4971..5106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5155..5949
                     /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             5155..5949
                     /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(5209..5228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5819..5838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6137..6156)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6181..6228
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7046..7065
                     /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 atcctgccat gggcaaccgc ggcatggaag atctcatccc gctggtcaac
      601 cggctgcaag acgccttctc tgccatcggc cagaacgcgg acctcgacct gccgcagatc
      661 gctgtggtgg gcggccagag cgccggcgcg agctcggtgc tcgagaattt cgtaggcagg
      721 gacttcttgc ctcgaggatc tggcattgtt acccgacgtc ccctggtctt gcagctggtc
      781 aatgcaacca cagaatatgc cgagttcctg cactgcaagg gaaagaaatt caccgacttc
      841 gaggaggtgc gccttgagat cgaggccgag accgacaggg tcaccggcac caacaagggc
      901 atctcgccgg tgcctatcaa cctccgcgtc tactcgccgc acgtgctgaa cctgaccctg
      961 gtggacctgc ccggaatgac caaggtcccg gtgggggacc aacctcccga catcgagttc
     1021 cagatccgag acatgcttat gcagtttgtc accaaggaga actgcctcat cctggccgtg
     1081 tcccccgcca actctgacct ggccaattct gacgccctca aggtcgccaa ggaggtggac
     1141 ccccagggcc agcgcaccat cggggtcatc accaagctgg acctgatgga cgagggcaca
     1201 gatgcccgtg atgtgctgga gaacaagctg ctccccctgc gcagaggcta cattggagtg
     1261 gtgaaccgga gccagaagga cattgatggc aagaaggaca ttaccgccgc cttggctgct
     1321 gaacgaaagt tcttcctctc ccatccatct tatcgccact tggctgaccg tatgggcacg
     1381 ccctacctgc agaaggtcct caatcagcaa ctgacgaacc acatccggga cacactgccg
     1441 gggctgcgga acaagctgca gagccagcta ctgtccattg agaaggaggt ggaggaatac
     1501 aagaacttcc gccctgatga cccagctcgc aagaccaagg ccctgctgca gatggtccag
     1561 cagtttgccg tagactttga gaagcgcatt gagggctcag gagatcagat cgacacctac
     1621 gaactgtcag ggggagcccg cattaaccga atcttccacg agcgcttccc tttcgagctg
     1681 gtcaagatgg agtttgatga gaaggaactc cgaagggaga tcagctatgc tatcaagaat
     1741 atccatggca ttagaacggg gctgtttacc ccagacatgg cctttgagac cattgtgaaa
     1801 aagcaggtga agaagatccg agaaccgtgt ctcaagtgtg tggacatggt tatctcggag
     1861 ctaatcagca ccgttagaca gtgcaccaag aagctccagc agtacccgcg gctacgggag
     1921 gagatggagc gcatcgtgac cacccacatc cgggagcgcg agggccgcac taaggagcag
     1981 gtcatgcttc tcatcgatat cgagctggct tacatgaaca ccaaccatga ggacttcata
     2041 ggctttgcca atgctcagca gaggagcaac cagatgaaca agaagaagac ttcagggaac
     2101 caggatgaga ttctggtcat ccgcaagggc tggctgacta tcaataatat tggcatcatg
     2161 aaagggggct ccaaggagta ctggtttgtg ctgactgctg agaatctgtc ctggtacaag
     2221 gatgatgagg agaaagagaa gaaatacatg ctgtctgtgg acaacctcaa gctgcgggac
     2281 gtggagaagg gctttatgtc gagcaagcat atctttgccc tctttaacac ggagcagagg
     2341 aatgtctaca aggattatcg gcagctggag ctagcctgtg agacacagga ggaggtggac
     2401 agctggaagg cctccttcct gagggctggc gtgtaccctg agcgtgttgg ggacaaagag
     2461 aaagccagcg agaccgagga gaatggctcc gacagcttca tgcattccat ggacccacag
     2521 ctggaacggc aagtggagac catccggaat cttgtggact catacatggc cattgtcaac
     2581 aagaccgtga gggacctcat gcccaagacc atcatgcacc tcatgattaa caataccaag
     2641 gagttcatct tctcggagct gctggccaac ctgtactcgt gtggggacca gaacacgctg
     2701 atggaggagt cggcggagca ggcacagcgg cgcgacgaga tgctgcgcat gtaccacgca
     2761 ctgaaggagg cgctcagcat catcggcaac atcaacacga ccaccgtcag cacgcccatg
     2821 cccccgcccg tggacgactc ctggctgcag gtgcagagcg taccggccgg acgcaggtcg
     2881 cccacgtcca gccccacgcc gcagcgccga gcccccgccg tgcccccagc ccggcccggg
     2941 tcgcggggcc ctgctcctgg gcctccgcct gctgggtccg ccctgggggg ggcgcccccc
     3001 gtgccctcca ggccgggggc ttcccctgac cctttcggcc ctccccctca ggtgccctcg
     3061 cgccccaacc gcgccccgcc cggggtcccc agccgatcgg gtcaggcaag tccatcccgt
     3121 cctgagagcc ccaggccccc cttcgacaag cttcgaattc tgcagtcgac ggtaccgcgg
     3181 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     3241 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3301 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3361 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3421 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3481 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3541 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3601 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3661 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3721 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3781 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3841 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3901 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     3961 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4021 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4081 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4141 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4201 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4261 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4321 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4381 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4441 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4501 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4561 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4621 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4801 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4861 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4921 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4981 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5041 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5101 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5161 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5221 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5281 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5341 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5401 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5461 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5521 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5581 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5641 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5701 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5761 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5821 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5881 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5941 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6001 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6061 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6121 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6181 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6241 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6301 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6361 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     7261 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
