LOCUS       sequence-358082-        5872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNA targeting mtIF3 mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5872)
  AUTHORS   Lee S, Park D, Lim C, Kim JI, Min KT
  TITLE     mtIF3 is locally translated in axons and regulates mitochondrial 
            translation for axonal growth.
  JOURNAL   BMC Biol. 2022 Jan 7;20(1):12. doi: 10.1186/s12915-021-01215-w.
  PUBMED    34996455
REFERENCE   2  (bases 1 to 5872)
  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: "BMC Biol.";
            date: "2022-01-7"; volume: "20(1):12. doi"; pages: " 
            10.1186/s12915-021-01215-w"
COMMENT     Sequence Label: pAAV2-mtIF3 shRNA_TagRFP657
FEATURES             Location/Qualifiers
     source          1..5872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             825..1526
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="MSELITENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSHTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEAHTEMLYPADGGLEGRTALALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDYRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    1670..2258
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1723..1743)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2141..2152)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2290..2766
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2489..2508)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2806..2946
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2806..2935
                     /label=AAV2 ITR
     rep_origin      3021..3476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3108..3127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3318..3339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3493..3512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3612..3634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3672..3690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3758..3862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3863..3931,3932..4723)
                     /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(4081..4100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4894..5482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5383..5402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5544..5673
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaggatccc cacgttcaag tcacgataaa gaagcttgtt tatcgtgact tgaacgtggt
      301 ttttttctag agggatcatc taatcaagca caaataaggg gcgtgtaaca caaaagccag
      361 cgaccctttc caatgcaaat caaacttgca attctttgcc gtttttatca tttaagtgtc
      421 ggcttaaggt ccactatcag atgtaaacag ccttatctaa caaaggtatc attacattct
      481 gaaattctca ggcatgcaag ctagcttatg acgcactagg gagtgccacc cttcctttcg
      541 ccctaacttc gtgataactc gcgcgtttca ctcaacagct gcatccgccc tagtgctact
      601 gggagttgta gtatacaaga cgcttacagg ctgaatgttc tgtcaagacc ccgcctctag
      661 cactttggga attctggact tgatgatgtc atggttaatc cccgcccagt agaggcggct
      721 atataaaggg tggttaaggc ccggttcgct ctcttcctca ccgggtctgc ggcgagttct
      781 agctgtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatgagc gagctgatca
      841 ccgagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt
      901 gcacatccga gggcgaaggc aagccctacg agggcaccca gacccagaga atcaaggtgg
      961 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
     1021 gtcacacctt catcaaccac acccagggca tccccgactt ctggaagcag tccttccctg
     1081 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     1141 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     1201 tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc cacaccgaga
     1261 tgctgtaccc cgctgacggc ggcctggaag gcagaaccgc gctggccctg aagctcgtgg
     1321 gcgggggcca cctgatctgc aacttcaaga ccacatacag atccaagaaa cccgctaaga
     1381 acctcaagat gcccggcgtc tactatgtgg actacagact ggaaagaatc aaggaggccg
     1441 acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     1501 gcaaactggg gcacaagctt aattaaagcg gccgcgactc tagatcataa tcagccatac
     1561 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1621 acataaaatg aatgcaattg ttgttgttat catcatcaag cttatcgata atcaacctct
     1681 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1741 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1801 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     1861 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     1921 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     1981 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2041 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     2101 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2161 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2221 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     2281 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     2341 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     2401 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     2461 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2521 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2581 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2641 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     2701 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     2761 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2821 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2881 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2941 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3001 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3061 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3121 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3181 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3241 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3301 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     3361 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     3421 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3481 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3541 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3601 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3661 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3721 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3781 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3841 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3901 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3961 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4021 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4081 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4141 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4201 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4261 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4321 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4381 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4441 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4501 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4561 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4621 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4681 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4741 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4801 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5041 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5521 ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc
     5581 cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     5641 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtaag ctttgcaaag
     5701 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
     5761 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     5821 cgagaagttg gggggagggg tcggcaattg aaccggtaga tctgggcagg aa
//
