LOCUS       sequence-379584-        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a Fis1-GFP chimera lacking the C-terminal 27 residues 
            (Fis1-DTM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5083)
  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            "unpublished"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: pEGFP-Fis1-DTM
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     586..605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(904..923)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    948..995
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1324..1912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1813..1832
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2087..2390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2391..2594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2545..2565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3049..3058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3055..3057,3058..3060,3061..3774)
                     /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(3055..3057,3058..3060,3061..3771)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3100..3121)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3361..3380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3708..3729
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3897..4018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3934..3953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3988..4007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4025..4480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4162..4183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4374..4393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4507..4611
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4613..4970
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4613..4804
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4636..4656)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4821..4956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5005..5083,1..716)
                     /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             join(5005..5083,1..716)
                     /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(5059..5078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
       61 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
      121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
      421 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
      481 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
      541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
      601 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
      661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
      721 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
      781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
      841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
      901 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
      961 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     1021 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     1081 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     1141 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     1201 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1261 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1441 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1681 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1741 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1801 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1861 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1921 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1981 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     2041 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2101 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2161 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2221 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2281 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2341 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2401 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2461 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2521 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2581 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
     2641 cgagctcaag cttcgaattc caccatggag gccgtgctga acgagctggt gtctgtggag
     2701 gacctgctga agtttgaaaa gaaatttcag tctgagaagg cagcaggctc ggtgtccaag
     2761 agcacgcagt ttgagtacgc ctggtgcctg gtgcggagca agtacaatga tgacatccgt
     2821 aaaggcatcg tgctgctcga ggagctgctg cccaaaggga gcaaggagga acagcgggat
     2881 tacgtcttct acctggccgt ggggaactac cggctcaagg aatacgagaa ggccttaaag
     2941 tacgtccgcg ggttgctgca gacagagccc cagaacaacc aggccaagga actggagcgg
     3001 ctcattgaca aggccatgaa gaaagatgga ctcgcggatc caccggtcgc caccatggtg
     3061 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3121 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3181 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3241 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     3301 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3361 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3421 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3481 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3541 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3601 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3661 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3721 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     3781 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3841 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3901 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3961 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4021 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4081 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4141 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4201 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4261 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4321 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4381 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4441 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4501 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4561 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4621 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4681 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4741 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4801 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4861 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4921 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4981 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5041 gccgcttggg tggagaggct attcggctat gactgggcac aac
//
