LOCUS       sequence-97686-e        7104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to make mCherry-Parkin stable cell line with retrovirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Yamano K, Fogel AI, Wang C, van der Bliek AM, Youle RJ
  TITLE     Mitochondrial Rab GAPs govern autophagosome biogenesis during 
            mitophagy.
  JOURNAL   Elife (Cambridge). 2014 Feb 25;3:e01612. doi: 10.7554/eLife.01612.
  PUBMED    24569479
REFERENCE   2  (bases 1 to 7104)
  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: "Elife 
            (Cambridge)."; date: "2014-02-25"; volume: "3:e01612. doi"; pages: "
            10.7554/eLife.01612"
COMMENT     Sequence Label: pBMN-mCherry-Parkin
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             541..1010
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1075..1432
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1497..1913
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1843..1865
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1881..1897
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1923..2297
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2235..2251
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      2306..2315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2312..3019
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2457..2475)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2725..2744
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4511..5104
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5235..5252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5406..5994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5486..5505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6168..6959,6960..7028))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6791..6810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     join(7096..7104,1..10)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atcacgaggc cctttcgtct tcaagaacag ctttgctctt aggagtttcc taatacatcc
       61 caaactcaaa tatataaagc atttgacttg ttctatgccc tagggggcgg ggggaagcta
      121 agccagcttt ttttaacatt taaaatgtta attccatttt aaatgcacag atgtttttat
      181 ttcataaggg tttcaatgtg catgaatgct gcaatattcc tgttaccaaa gctagtataa
      241 ataaaaatag ataaacgtgg aaattactta gagtttctgt cattaacgtt tccttcctca
      301 gttgacaaca taaatgcgct gctgagaagc cagtttgcat ctgtcaggat caatttccca
      361 ttatgccagt catattaatt actagtcaat tagttgattt ttatttttga catatacatg
      421 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
      481 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag
      541 ctgaatatgg gccaaagcgg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      601 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      661 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      721 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      781 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      841 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      901 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      961 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
     1021 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
     1081 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
     1141 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
     1201 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg
     1261 tggcccgacc tgagtccaaa aatcccgatc gttttggact ctttggtgca ccccccttag
     1321 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     1381 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
     1441 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaatatg ggcccgggcc
     1501 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1561 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1621 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1681 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1741 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1801 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1861 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
     1921 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
     1981 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
     2041 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     2101 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     2161 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     2221 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     2281 gaccatcctc tagactgccg gatccgccac catggtgagc aagggcgagg aggataacat
     2341 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     2401 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2461 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2521 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2581 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2641 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2701 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2761 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2821 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2881 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2941 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     3001 catggacgag ctgtacaagt ccggactcag atctcgagct caagctttga tagtgtttgt
     3061 caggttcaac tccagccatg gtttcccagt ggaggtcgat tctgacacca gcatcttcca
     3121 gctcaaggag gtggttgcta agcgacaggg ggttccggct gaccagttgc gtgtgatttt
     3181 cgcagggaag gagctgagga atgactggac tgtgcagaat tgtgacctgg atcagcagag
     3241 cattgttcac attgtgcaga gaccgtggag aaaaggtcaa gaaatgaatg caactggagg
     3301 cgacgacccc agaaacgcgg cgggaggctg tgagcgggag ccccagagct tgactcgggt
     3361 ggacctcagc agctcagtcc tcccaggaga ctctgtgggg ctggctgtca ttctgcacac
     3421 tgacagcagg aaggactcac caccagctgg aagtccagca ggtagatcaa tctacaacag
     3481 cttttatgtg tattgcaaag gcccctgtca aagagtgcag ccgggaaaac tcagggtaca
     3541 gtgcagcacc tgcaggcagg caacgctcac cttgacccag ggtccatctt gctgggatga
     3601 tgttttaatt ccaaaccgga tgagtggtga atgccaatcc ccacactgcc ctgggactag
     3661 tgcagaattt ttctttaaat gtggagcaca ccccacctct gacaaggaaa catcagtagc
     3721 tttgcacctg atcgcaacaa atagtcggaa catcacttgc attacgtgca cagacgtcag
     3781 gagccccgtc ctggttttcc agtgcaactc ccgccacgtg atttgcttag actgtttcca
     3841 cttatactgt gtgacaagac tcaatgatcg gcagtttgtc cacgaccctc agcttggcta
     3901 ctccctgcct tgtgtggctg gctgtcccaa ctccttgatt aaagagctcc atcacttcag
     3961 gattctggga gaagagcagt acaaccggta ccagcagtat ggtgcagagg agtgtgtcct
     4021 gcagatgggg ggcgtgttat gcccccgccc tggctgtgga gcggggctgc tgccggagcc
     4081 tgaccagagg aaagtcacct gcgaaggggg caatggcctg ggctgtgggt ttgccttctg
     4141 ccgggaatgt aaagaagcgt accatgaagg ggagtgcagt gccgtatttg aagcctcagg
     4201 aacaactact caggcctaca gagtcgatga aagagccgcc gagcaggctc gttgggaagc
     4261 agcctccaaa gaaaccatca agaaaaccac caagccctgt ccccgctgcc atgtaccagt
     4321 ggaaaaaaat ggaggctgca tgcacatgaa gtgtccgcag ccccagtgca ggctcgagtg
     4381 gtgctggaac tgtggctgcg agtggaaccg cgtctgcatg ggggaccact ggttcgacgt
     4441 gtaggcggcc gccagcacag tggtcgacga taaaataaaa gattttattt agtctccaga
     4501 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     4561 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     4621 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     4681 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     4741 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     4801 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     4861 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     4921 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     4981 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     5041 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     5101 cattcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa
     5161 atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5221 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5281 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5341 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5821 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6061 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6121 ttgcgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6181 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6241 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6301 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6361 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6421 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6481 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6541 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6601 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6661 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6721 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6781 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6841 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6901 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6961 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7021 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7081 acattaacct ataaaaatag gcgt
//
