LOCUS       sequence-199732-        7373 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 7373)
  TITLE     wt HA-dynamin 1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7373)
  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: 
            "Unpublished"
COMMENT     Sequence Label: K44A dynamin 2 pEGFP
FEATURES             Location/Qualifiers
     source          1..7373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1369..1380
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3325..3334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3331..3333,3334..3336,3337..4050)
                     /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(3331..3333,3334..3336,3337..4047)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3376..3397)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3637..3656)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3984..4005
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4173..4294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4210..4229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4264..4283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4301..4756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4438..4459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4650..4669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4783..4887
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4889..5246
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4889..5080
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4912..4932)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5097..5232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5281..6075
                     /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             5281..6075
                     /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(5335..5354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5945..5964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6263..6282)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6307..6354
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6683..7271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7172..7191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcgc taccggactc agatctcgag ctcaagcttg gcaccatggg caaccgcggg
      661 atggaagagc tcatcccgct ggttaacaag ctgcaggacg ccttcagctc catcggtcag
      721 agctgccacc tggacctgcc gcagatcgcc gtggtgggcg gccagagcgc cggcgcgagt
      781 tcggtgctcg agaacttcgt gggccgggac ttccttccac gaggatcagg aattgtcacc
      841 cggaggcctc tcattctgca gctcatcttt tccaaaacag aatatgcgga gtttttgcac
      901 tgcaagtcca aaaagtttac agactttgat gaagtccggc aggagatcga agcagagact
      961 gaccgggtca caggcaccaa caaaggcatc tccccggtgc ccatcaacct tcgggtctat
     1021 tcaccacacg tgttgaactt gaccctcatc gacctcccag gcatcactaa ggtgccagtg
     1081 ggggaccagc caccagacat cgagtaccag atcaaggaca tgatcctgca gtttatcagc
     1141 cgggagagca gcctcattct tgccgtcaca cctgccaaca tggacttggc caactcggat
     1201 gccctcaagc tggccaagga ggtggacccc caaggcctgc ggaccattgg tgtcatcacc
     1261 aagctagacc tgatggatga aggcacagat gccagggatg tcctggagaa caagctcctg
     1321 cccttgagaa gaggctacat cggcgtggtt aaccgcagcc agaaagacat cgagggcaga
     1381 aaggacatcc gggcagctct ggcagctgag aggaaattct tcctctccca cccagcctac
     1441 cggcacatgg ctgaccgcat gggtacccca cacttgcaga aaacactgaa ccagcaactg
     1501 accaaccaca tccgggagtc actgccaacc ctgcgcagca agctgcagag ccaactgctg
     1561 tccctggaga aggaagtgga agagtacaag aatttccggc ctgacgaccc cacacgcaag
     1621 accaaagccc tgctgcagat ggtccagcag tttggagtgg actttgagaa gcgaatcgaa
     1681 ggctctggag atcaagtcga caccctggag ctgtctgggg gcgcccgcat caatcgcatc
     1741 ttccacgagc gctttccctt tgagctggtg aagatggagt ttgacgagaa agacctccga
     1801 agagagatca gctacgccat taagaacatc cacggagtca ggacggggct cttcaccccc
     1861 gacatggcct ttgaagccat cgtgaaaaaa caacttgtaa agctcaaaga gccgagtttg
     1921 aagtgtgttg acctggtggt gtcggagcta gccacggtca ttaaaaagtg tgctgagaag
     1981 ctcagttcct acccccgcct tcgagaggag accgaacgca ttgtcaccac ctacatcagg
     2041 gagcgagaag ggagaaccaa ggaccagatc cttctcctga ttgacattga gcagtcctac
     2101 atcaacacaa accatgaaga cttcattgga tttgccaatg cccagcagag gagcacgcag
     2161 ctgaacaaga agagggccat acctaaccag ggggagatct tggtgatccg caggggctgg
     2221 ttgaccatca acaacatcag cttgatgaaa ggcggttcca aggagtactg gttcgtgctg
     2281 actgctgagt cgttgtcttg gtacaaggat gaagaggaaa aagaaaagaa gtacatgctg
     2341 ccactagaca acctcaaaat acgggatgtg gagaagggct tcatgtccaa caagcatgtg
     2401 tttgccatct tcaacacaga gcagaggaac gtctacaagg acctgcgaca gatcgaactg
     2461 gcttgtgact cccaggaaga tgtggacagc tggaaggctt cgttcctgcg agctggggtc
     2521 tacccagaga aggaccaggc agagaacgag gatggagcac aagagaacac cttctccatg
     2581 gacccgcagc tggagcggca ggtggagacc atccgcaact tggtggactc ctatgtggcc
     2641 atcatcaaca agtccatccg ggacctcatg ccaaagacca tcatgcacct catgatcaac
     2701 aacacaaagg ccttcatcca ccatgagctg ctagcctacc tgtactcatc agcagaccag
     2761 agcagcctca tggaggagtc agctgagcag gctcagcggc gggacgatat gctgcgcatg
     2821 taccatgcac tcaaggaagc gctcaacatc attggggaca tcagcaccag caccgtgtcc
     2881 acgcctgtgc ccccacctgt cgatgatact tggctccaga acaccagcag ccacagcccc
     2941 actccacagc gcagacctgt gtccagcgtg cacccgccgg gccgcccacc cgcagtgcgg
     3001 ggccccacac cggggcctcc cctcattcct atgcctgtgg gggccacatc ctccttctcg
     3061 gcacccccca tcccatcccg gcctggacca cagaatgtgt ttgccaacaa tgaccccttc
     3121 tctgcgccac ctcagatacc gtctcggcca gcacggattc cccctgggat ccctccagga
     3181 gtgcccagca gaagagcgcc tgctgcgccc agccggccca ccattatccg cccagccgag
     3241 ccgtccctgc tcgacgctgc aggggacagt gttctggggg gaattctgca gtcgacggta
     3301 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     3361 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3421 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3481 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3541 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3601 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3661 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3721 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3781 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3841 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3901 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     3961 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4021 actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc
     4081 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4141 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4201 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4261 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4321 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4381 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4441 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4501 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4561 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4621 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4681 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4741 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4861 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4921 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4981 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5041 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5101 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5161 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5221 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5281 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5341 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5401 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5461 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5521 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5581 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5641 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5701 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5761 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5821 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5881 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5941 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6001 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6061 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6121 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6181 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6241 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6301 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6361 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6421 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6481 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     6541 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6601 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6661 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6721 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6781 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6841 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6901 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6961 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7021 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7081 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7141 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7201 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7261 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7321 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt att
//
