LOCUS       sequence-379560-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a Drp1-ECFP chimera.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6804)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6804)
  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            "unpublished"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: pECFP-Drp1
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(226..583)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      240..375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        392..583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     540..560
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(585..689)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      716..1171
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(803..822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1013..1034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1178..1299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1189..1208)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1243..1262
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3562..4272,4273..4275,4276..4278))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3604..3625)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3953..3972
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4212..4233
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4275..4284
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4323..4526)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4352..4372)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4527..4830
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5005..5593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5085..5104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5922..5969
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5994..6013
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(6201..6804,1..191))
                     /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             complement(join(6201..6804,1..191))
                     /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(6312..6331)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
       61 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
      121 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
      181 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag
      241 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
      301 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
      361 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
      421 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
      481 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      541 ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa
      601 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
      661 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
      721 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      781 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      841 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
      901 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
      961 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1021 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1081 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1141 aattttaaca aaatattaac gcttacaatt tacgccttaa gatacattga tgagtttgga
     1201 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1261 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1321 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1381 aaatgtggta tggctgatta tgatcagtta tctagatccg gtggatcctc accaaagatg
     1441 agtctcccgg atttcagcaa taatttgact ggctccttgt aatgccttta gcatatcagc
     1501 tgcttctttc ctgcgctgtg ccatgtcctc agattctgtc agaagatcat ccaataagga
     1561 tgatttatac agctggccta ctagctcact ctgaagagtg tctttcacat gattaaccaa
     1621 aaaatgcatt actgcctttg gcacactgtc ttgaatattc tttctgacaa tgagaaaata
     1681 tgatttaatg agtcgttcaa taacctcaca atctcgctgt tcccgagcag atagttttcg
     1741 tgcaacagga actggcacat ctagcaggtt cacggcatga cctttttgtg gactggctgg
     1801 cataattgga atgggttttg atttttcttc tgctaataac tcttcagctt ttgaagtttt
     1861 cagcattcct ctccagttgc ctgtggttgg ttcttgaaca ccatctccaa ccccaccacc
     1921 tccagatgca accttgtctc gtgatacagc tgaaggtaat tctctggcta gcctgtttct
     1981 cctttgttcc tctatattat tgttcattag cccacaagca tcagcaaagt ctggatgttt
     2041 tgtgttgata taagccagtt caattgccac taagttatgg accatttcat ttgtaacagg
     2101 caacctttta cgaagaagac aagtcaccac ttcaactatg gcatcatgaa gtttaggaaa
     2161 tcgtaacaat tcctgtgtac tgtaattgct acagtgctga atgatccttt gcatttcctc
     2221 atgaaccagt tccacacagc ggaggctggg ctcttctaga cgtttgattt gccgcttcac
     2281 cagtaactca aatgaaacct caggcacaaa taaagcagga cgaggaccag tagcatttct
     2341 aatggcagtc aaaatgtcaa tagtgttaag gccaccaagt ggatcaacag attctaaggt
     2401 tcgcccaaaa gtctcatgga aaatataaca aattctagca ccaccgcata gctccgaagt
     2461 ttcaatatat tttgcagttc cttcaatagt gttacaatat tctgtggcaa atttggtaat
     2521 aagttggagt aaagtagcac ttttatcatc cacgggttca ccgtagctat ttagaagaga
     2581 ctgatactga gcagctagaa catttattct tgttttcaac tctggtaaac aatctctgat
     2641 gtgatgcatc agtaacctgt ttagagtcct agcaagatac tttgttccat ttctattggc
     2701 cagagatgga tatttctttt gaagaaaagc atactcatca cggattgaat cagttacact
     2761 cttcttgttg ttaatatcta gctggctcct gttaactact ccaattattc caagtttgac
     2821 tggaataacc cttcccatca atacatccat ggcatcagta cccgcatcca tgagatcaag
     2881 tttagtgatt acagctaggg ttctgcgacc atctggatct acctctcttg aaattttaag
     2941 tgcctctgat gttgccatat ctgtattagc agcagtgaca gcgaggataa tggaattagg
     3001 attactgatg aaccgaagaa tgagctctct gatttgaagc tcaatatcct taggttgatc
     3061 acctacaggc accttggtca ttcctggcaa atccacaagt gtcaaattga caacgttggg
     3121 tgaaaaaatc ttaagatgaa ttggttcagg gcttactccc ttattatttc ctgaaattct
     3181 ttctgtttca ttttcaattt cttgtcgaat ttcatcaaaa tccgtgtaaa gcttattttt
     3241 ggtgtgaaga aatttacccc attcttctgc ttccacccca ttttcttctc ctgttgtttt
     3301 ccgtttatct tcttgtgaaa catggaccag ttgcagaatg agaggtctcc gggtgacaat
     3361 tccagtacct ctgggaagca ggtccctccc caccaggctt tctagcactg agctcgctcc
     3421 gctgctctgc gttcccacta cgacgatttg aggcagctgg atgatgtcgg cgcccaccgt
     3481 gttgaagacg tcctggagct tgtttatgac aggaattagc gcctcagaat tcgaagcttg
     3541 agctcgagat ctgagtccgg acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     3601 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     3661 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     3721 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     3781 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgat
     3841 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     3901 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     3961 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     4021 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgcccca
     4081 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     4141 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     4201 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     4261 ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ggatctgacg gttcactaaa
     4321 ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     4381 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     4441 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     4501 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     4561 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     4621 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     4681 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     4741 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     4801 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     4861 gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga
     4921 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4981 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5041 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5101 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5161 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5221 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5281 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5341 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtga
     5401 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5461 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5521 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5581 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5641 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5701 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     5761 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     5821 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     5881 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     5941 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     6001 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     6061 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     6121 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     6181 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     6241 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     6301 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     6361 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     6421 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     6481 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     6541 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     6601 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     6661 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     6721 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     6781 gtggccagcc acgatagccg cgct
//
