LOCUS       sequence-192863-        6376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses shRNA against human Drp1 from puromycin resistance 
            retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  AUTHORS   Sweatt AJ, Griffiths CD, Groves SM, Paudel BB, Wang L, Kashatus DF, 
            Janes KA
  TITLE     Proteome-wide copy-number estimation from transcriptomics.
  JOURNAL   Mol Syst Biol. 2024 Sep 27. doi: 10.1038/s44320-024-00064-3.
  PUBMED    39333715
REFERENCE   2  (bases 1 to 6376)
  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: "Mol Syst 
            Biol. 2024 Sep 27. doi: 10.1038/s44320-024-00064-3."
COMMENT     Sequence Label: pSuper-Retro-Puro-Drp1-shRNA
FEATURES             Location/Qualifiers
     source          1..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(288..304)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(289..305)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(323..739)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(339..355)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(371..393)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    806..1147
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             1211..1727
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1927..1946)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2074..2096)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2290..2309)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2409..2431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2469..2487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2555..2659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2660..2728,2729..3520)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2878..2897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3691..4279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4180..4199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4433..4450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4567..4588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4603..4608,4609..4626,4627..4633)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4641..4657
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             complement(5243..5842)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5326..5346)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5823..5842
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5867..6366)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5915..5934)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6308..6331
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacagatc ccccaggagc cagctagata
      241 ttaattcaag agattaatat ctagctggct cctgttttta agcttatcga taccgtcgac
      301 ctcgagagat cgatctaatt ccggcgccta gagaaggagt gagggctgga taaagggagg
      361 atcgaggcgg ggtcgaacga ggaggttcaa gggggagaga cggggcggat ggaggaagag
      421 gaggcggagg cttagggtgt acaaagggct tgacccaggg aggggggtca aaagccaagg
      481 cttcccaggt cacgatgtag gggacctggt ctgggtgtcc atgcgggcca ggtgaaaaga
      541 ccttgatctt aacctgggtg atgaggtctc ggttaaaggt gccgtctcgc ggccatccga
      601 cgttaaaggt tggccattct gcagagcaga aggtaaccca acgtctcttc ttgacatcta
      661 ccgactggtt gtgagcgatc cgctcgacat ctttccagtg acctaaggtc aaacttaagg
      721 gagtggtaac agtctggccc taattttcag acaaatacag aaacacagtc agacagagac
      781 aacacagaac gatgctgcag cgctgcagca gacaagacgc gcggcttcgg ttccaaaccg
      841 aaagcaaaaa ttcagacgga ggcgggaact gttttaggtt ctcgtctcct accagaacca
      901 catatcctga cggggtcgga ttccacatcg actcccttcc tcaggtcggg ccacaaaaac
      961 ggcccccaaa gtccctggga cgtctcccag ggttgcggcc gggtgttcag aactcgtcag
     1021 ttccaccacg ggtccgccag atacagagct agttagctaa ctagtacaga cgcaggcgca
     1081 aacattagat gccggcacaa acacgcacaa agacagagac agacacgaaa cgaccgctgg
     1141 ccagcttacc tcccggcggg ggggtcggtg gtccctgggc aggggtctcc aaatctcggt
     1201 ggaacctcca aatgaaagac ccccgaggtg ggcagtcaat caatctgagg agaccctccc
     1261 aaggatcagc gagtccacga ttcggatgca aacagcaaga ggctttattg ggaatacggg
     1321 tacccgggcg acgcagtcta tcggaggact ggcgcgccga gtgaggggtt gtgggctctt
     1381 ttattgagct cggggagcag aagcgcgcga acagaagcga gaagcgaact gattggttag
     1441 ttcaaataag gcacagggtc atttcaggtc cttggggcac cctggaaaca tctgatggtt
     1501 ctctagaaac tgctgagggc gggaccgcat ctggggacca tctgttcttg gccctgagcc
     1561 ggggcaggaa ctgcttacca cagatatcct gtttggccca tattctgctg tctctctgtt
     1621 cctaaccttg atctgaactt ctctattctc agttatgtat tttccatgcc ttgcaaaatg
     1681 gcgttactta agctagcttg ccaaacctac aggtggggtc tttcattccc ccctttttct
     1741 ggagactaaa taaaatcttt tattttatct atggctcgta ctctataggc ttcagctggt
     1801 gatattgttg agtcaaaact agagcctgga ccactgatat cctgtcttta acaaattgga
     1861 ctaatcgcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     1921 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     1981 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     2041 ccatctcctt gcatgcacca ttccttgcgc cgtcgtttta caacgtcgtg actgggaaaa
     2101 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2161 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2221 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     2281 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2341 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2401 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2461 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2521 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2581 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2641 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2761 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2821 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2941 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3061 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3241 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3361 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3541 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3601 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4261 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4321 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4381 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4441 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4501 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4561 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4621 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacaattctc cccagctctt
     4681 tctgcgctgt ccataatgaa ctatttcagg agtattggag tatgtggatt caaaacagtg
     4741 gctaattgag tcaggattac ttaatattcg actcctaccc tcagaatgtt gttctccaac
     4801 ttcaagaaac tgttacccat gaaagacccc cgctgacggg tagtcaatca ctcagaggag
     4861 accctcccaa ggaacagcga gaccacaagt cggatgcaac tgcaagaggg tttattggat
     4921 acacgggtac ccgggcgacg cagtctatcg gaggactggc gcgccgagtg aggggttgtg
     4981 ggctctttta ttgagctcgg ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat
     5041 tggttagttc aaataaggca cagggtcatt tcaggtcctt ggggcaccct ggaaacatct
     5101 gatggttctc tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga
     5161 gactaaataa aatcttttat tttatcgatg catggggtcg tgcgctcctt tcggtcgggc
     5221 gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
     5281 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
     5341 ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
     5401 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga
     5461 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
     5521 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
     5581 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
     5641 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
     5701 cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg
     5761 cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca
     5821 ccgtgggctt gtactcggtc atggtaagct agcttgggct gcaggtcgaa aggcccggag
     5881 atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc
     5941 ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg
     6001 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc
     6061 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag
     6121 tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga
     6181 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg
     6241 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc
     6301 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc
     6361 ctacccggta gaattc
//
