LOCUS       sequence-215520-        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   Lee E, De Camilli P
  TITLE     Dynamin at actin tails.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Jan 8. 99(1):161-6.
  PUBMED    11782545
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: "Proc Natl 
            Acad Sci U S A. 2002 Jan 8. 99(1):161-6."
COMMENT     Sequence Label: pEGFPN1-human Dynamin 1aa K44A
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3962..3971
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3968..3970,3971..3973,3974..4687)
                     /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(3968..3970,3971..3973,3974..4684)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4013..4034)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4274..4293)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4621..4642
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4810..4931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4847..4866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4901..4920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4938..5393)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5075..5096)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5287..5306
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5420..5524
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5526..5883
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5526..5717
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5549..5569)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5734..5869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5918..6712
                     /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             5918..6712
                     /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(5972..5991)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6582..6601
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6900..6919)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6944..6991
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatcctgc
     1321 catgggcaac cgcggcatgg aagatctcat cccgctggtc aaccggctgc aagacgcctt
     1381 ctctgccatc ggccagaacg cggacctcga cctgccgcag atcgctgtgg tgggcggcca
     1441 gagcgccggc gcgagctcgg tgctcgagaa tttcgtaggc agggacttct tgcctcgagg
     1501 atctggcatt gttacccgac gtcccctggt cttgcagctg gtcaatgcaa ccacagaata
     1561 tgccgagttc ctgcactgca agggaaagaa attcaccgac ttcgaggagg tgcgccttga
     1621 gatcgaggcc gagaccgaca gggtcaccgg caccaacaag ggcatctcgc cggtgcctat
     1681 caacctccgc gtctactcgc cgcacgtgct gaacctgacc ctggtggacc tgcccggaat
     1741 gaccaaggtc ccggtggggg accaacctcc cgacatcgag ttccagatcc gagacatgct
     1801 tatgcagttt gtcaccaagg agaactgcct catcctggcc gtgtcccccg ccaactctga
     1861 cctggccaat tctgacgccc tcaaggtcgc caaggaggtg gacccccagg gccagcgcac
     1921 catcggggtc atcaccaagc tggacctgat ggacgagggc acagatgccc gtgatgtgct
     1981 ggagaacaag ctgctccccc tgcgcagagg ctacattgga gtggtgaacc ggagccagaa
     2041 ggacattgat ggcaagaagg acattaccgc cgccttggct gctgaacgaa agttcttcct
     2101 ctcccatcca tcttatcgcc acttggctga ccgtatgggc acgccctacc tgcagaaggt
     2161 cctcaatcag caactgacga accacatccg ggacacactg ccggggctgc ggaacaagct
     2221 gcagagccag ctactgtcca ttgagaagga ggtggaggaa tacaagaact tccgccctga
     2281 tgacccagct cgcaagacca aggccctgct gcagatggtc cagcagtttg ccgtagactt
     2341 tgagaagcgc attgagggct caggagatca gatcgacacc tacgaactgt cagggggagc
     2401 ccgcattaac cgaatcttcc acgagcgctt ccctttcgag ctggtcaaga tggagtttga
     2461 tgagaaggaa ctccgaaggg agatcagcta tgctatcaag aatatccatg gcattagaac
     2521 ggggctgttt accccagaca tggcctttga gaccattgtg aaaaagcagg tgaagaagat
     2581 ccgagaaccg tgtctcaagt gtgtggacat ggttatctcg gagctaatca gcaccgttag
     2641 acagtgcacc aagaagctcc agcagtaccc gcggctacgg gaggagatgg agcgcatcgt
     2701 gaccacccac atccgggagc gcgagggccg cactaaggag caggtcatgc ttctcatcga
     2761 tatcgagctg gcttacatga acaccaacca tgaggacttc ataggctttg ccaatgctca
     2821 gcagaggagc aaccagatga acaagaagaa gacttcaggg aaccaggatg agattctggt
     2881 catccgcaag ggctggctga ctatcaataa tattggcatc atgaaagggg gctccaagga
     2941 gtactggttt gtgctgactg ctgagaatct gtcctggtac aaggatgatg aggagaaaga
     3001 gaagaaatac atgctgtctg tggacaacct caagctgcgg gacgtggaga agggctttat
     3061 gtcgagcaag catatctttg ccctctttaa cacggagcag aggaatgtct acaaggatta
     3121 tcggcagctg gagctagcct gtgagacaca ggaggaggtg gacagctgga aggcctcctt
     3181 cctgagggct ggcgtgtacc ctgagcgtgt tggggacaaa gagaaagcca gcgagaccga
     3241 ggagaatggc tccgacagct tcatgcattc catggaccca cagctggaac ggcaagtgga
     3301 gaccatccgg aatcttgtgg actcatacat ggccattgtc aacaagaccg tgagggacct
     3361 catgcccaag accatcatgc acctcatgat taacaatacc aaggagttca tcttctcgga
     3421 gctgctggcc aacctgtact cgtgtgggga ccagaacacg ctgatggagg agtcggcgga
     3481 gcaggcacag cggcgcgacg agatgctgcg catgtaccac gcactgaagg aggcgctcag
     3541 catcatcggc aacatcaaca cgaccaccgt cagcacgccc atgcccccgc ccgtggacga
     3601 ctcctggctg caggtgcaga gcgtaccggc cggacgcagg tcgcccacgt ccagccccac
     3661 gccgcagcgc cgagcccccg ccgtgccccc agcccggccc gggtcgcggg gccctgctcc
     3721 tgggcctccg cctgctgggt ccgccctggg gggggcgccc cccgtgccct ccaggccggg
     3781 ggcttcccct gaccctttcg gccctccccc tcaggtgccc tcgcgcccca accgcgcccc
     3841 gcccggggtc cccagccgat cgggtcaggc aagtccatcc cgtcctgaga gccccaggcc
     3901 ccccttcgac aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt
     3961 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4021 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4081 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     4141 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     4201 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     4261 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     4321 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     4381 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     4441 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     4501 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4561 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     4621 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4681 caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     4741 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4801 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4861 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4921 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4981 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5041 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5101 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5161 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5221 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     5281 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     5341 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     5401 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5521 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5581 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5641 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5701 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5761 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5821 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5881 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5941 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6001 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     6061 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     6121 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6181 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6241 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6301 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6361 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6421 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     6481 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6541 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6601 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6661 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6721 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6781 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6841 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     6901 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     6961 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     7021 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     7081 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     7141 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     7201 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7261 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
