LOCUS       sequence-137171-        6984 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 6984)
  AUTHORS   Karbowski M, Lee YJ, Gaume B, Jeong SY, Frank S, Nechushtan A, 
            Santel A, Fuller M, Smith CL, Youle RJ
  TITLE     Spatial and temporal association of Bax with mitochondrial fission 
            sites, Drp1, and Mfn2 during apoptosis.
  JOURNAL   J Cell Biol. 2002 Dec 23. 159(6):931-8.
  PUBMED    12499352
REFERENCE   2  (bases 1 to 6984)
  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: "J Cell 
            Biol. 2002 Dec 23. 159(6):931-8."
COMMENT     Sequence Label: Mfn2-YFP
FEATURES             Location/Qualifiers
     source          1..6984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             1562..1570
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      2931..2940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2937..2939,2940..2942,2943..3656)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2982..3003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3243..3262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3590..3611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3779..3900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3816..3835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3870..3889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3907..4362)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4044..4065)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4256..4275
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4389..4493
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4495..4852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4495..4686
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4518..4538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4703..4838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4887..5681
                     /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             4887..5681
                     /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(4941..4960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5551..5570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5869..5888)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5913..5960
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6289..6877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6778..6797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt cgcgatgtcc ctgctcttct
      661 ctcgatgcaa ctctatcgtc acagtcaaga aaaataagag acacatggct gaggtgaatg
      721 catccccact taagcacttt gtcactgcca agaagaagat caatggcatt tttgagcagc
      781 tgggggccta catccaggag agcgccacct tccttgaaga cacgtacagg aatgcagaac
      841 tggaccccgt taccacagaa gaacaggttc tggacgtcaa aggttaccta tccaaagtga
      901 gaggcatcag tgaggtgctg gctcggaggc acatgaaagt ggcttttttt ggccggacga
      961 gcaatgggaa gagcaccgtg atcaatgcca tgctctggga caaagttctg ccctctggga
     1021 ttggccacac caccaattgc ttcctgcggg tagagggcac agatggccat gaggcctttc
     1081 tccttaccga gggctcagag gaaaagagga gtgccaagac tgtgaaccag ctggcccatg
     1141 ccctccacca ggacaagcag ctccatgccg gcagcctagt gagtgtgatg tggcccaact
     1201 ctaagtgccc acttctgaag gatgacctcg ttttgatgga cagccctggt attgatgtca
     1261 ccacagagct ggacagctgg attgacaagt tttgtctgga tgctgatgtg tttgtgctgg
     1321 tggccaactc agagtccacc ctgatgcaga cggaaaagca cttcttccac aaggtgagtg
     1381 agcgtctctc ccggccaaac atcttcatcc tgaacaaccg ctgggatgca tctgcctcag
     1441 agcccgagta catggaggag gtgcggcggc agcacatgga gcgttgtacc agcttcctgg
     1501 tggatgagct gggcgtggtg gatcgatccc aggccgggga ccgcatcttc tttgtgtctg
     1561 ctaaggaggt gctcaacgcc aggattcaga aagcccaggg catgcctgaa ggagggggcg
     1621 ctctcgcaga aggctttcaa gtgaggatgt ttgagtttca gaattttgag aggagatttg
     1681 aggagtgcat ctcccagtct gcagtgaaga ccaagtttga gcagcacacg gtccgggcca
     1741 agcagattgc agaggcggtt cgactcatca tggactccct gcacatggcg gctcgggagc
     1801 agcaggttta ctgcgaggaa atgcgtgaag agcggcaaga ccgactgaaa tttattgaca
     1861 aacagctgga gctcttggct caagactata agctgcgaat taagcagatt acggaggaag
     1921 tggagaggca ggtgtcgact gcaatggccg aggagatcag gcgcctctct gtactggtgg
     1981 acgattacca gatggacttc cacccttctc cagtagtcct caaggtttat aagaatgagc
     2041 tgcaccgcca catagaggaa ggactgggtc gaaacatgtc tgaccgctgc tccacggcca
     2101 tcaccaactc cctgcagacc atgcagcagg acatgataga tggcttgaaa cccctccttc
     2161 ctgtgtccgt gcggagtcag atagacatgc tggtcccacg ccagtgcttc tccctcaact
     2221 atgacctaaa ctgtgacaag ctgtgtgctg acttccagga agacattgag ttccatttct
     2281 ctctcggatg gaccatgctg gtgaataggt tcctgggccc caagaacagc cgtcgggcct
     2341 tgatgggcta caatgaccag gtccagcgtc ccatccctct gacgccagcc aaccccagca
     2401 tgcccccact gccacagggc tcgctcaccc aggaggagtt catggtttcc atggttaccg
     2461 gcctggcctc cttgacatcc aggacctcca tgggcattct tgttgttgga ggagtggtgt
     2521 ggaaggcagt gggctggcgg ctcattgccc tctcctttgg gctctatggc ctcctctacg
     2581 tctatgagcg tctgacctgg accaccaagg ccaaggagag ggccttcaag cgccagtttg
     2641 tggagcatgc cagcgagaag ctgcagcttg tcatcagcta cactggctcc aactgcagcc
     2701 accaagtcca gcaggaactg tctgggacct ttgctcatct gtgtcagcaa gttgacgtca
     2761 cccgggagaa cctggagcag gaaattgccg ccatgaacaa gaaaattgag gttcttgact
     2821 cacttcagag caaagcaaag ctgctcagga ataaagccgg ttggttggac agtgagctca
     2881 acatgttcac acaccagtac ctgcagccca gcagagcgga tccaccggtc gccaccatgg
     2941 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3001 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3061 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3121 tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     3181 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3241 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3301 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3361 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3421 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3481 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3541 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3601 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     3661 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3721 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3901 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3961 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4021 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4081 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4141 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4201 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4261 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4321 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4441 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4501 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4561 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4681 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4741 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4801 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4861 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4921 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4981 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5041 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5101 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5161 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5221 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5281 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5341 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5401 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5461 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5521 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5581 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5641 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5701 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5761 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5821 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5881 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5941 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6001 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6061 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6121 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6181 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6241 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6421 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6961 ttctgtggat aaccgtatta ccgc
//
