LOCUS       sequence-346729-        4727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSMART Tmem192-3X HA (targeting vector for genomic tagging).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4727)
  AUTHORS   Eapen VV, Swarup S, Hoyer MJ, Paulo JA, Harper JW
  TITLE     Quantitative proteomics reveals the selectivity of ubiquitin-binding
            autophagy receptors in the turnover of damaged lysosomes by 
            lysophagy.
  JOURNAL   eLife. 2021 Sep 29;10. pii: 72328. doi: 10.7554/eLife.72328.
  PUBMED    34585663
REFERENCE   2  (bases 1 to 4727)
  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."; 
            date: "2021-09-29"; volume: "10. pii: 72328. doi"; pages: " 
            10.7554/eLife.72328"
COMMENT     Sequence Label: pSMART Tmem192-3X HA (targeting vector for gen...
FEATURES             Location/Qualifiers
     source          1..4727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    731..764
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      1298..1329
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1330..1432
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(1433..1501,1502..2293)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1651..1670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2317..2344
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2356..2943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2436..2455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2965..2994)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             3566..3592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3605..3631
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3644..3670
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3716..3749
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3845..3868)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4195..4214
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(4302..4727,1..174)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4302..4321)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
       61 tccgcgcccc acaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
      121 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
      181 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gatgatatca
      241 gatccccggg atgcagaaat tgatgatcta ttaaacaata aagatgtcca ctaaaatgga
      301 agtttttcct gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg
      361 aaggattgga gctacggggg tgggggtggg gtgggattag ataaatgcct gctctttact
      421 gaaggctctt tactattgct ttatgataat gtttcatagt tggatatcat aatttaaaca
      481 agcaaaacca aattaagggc cagctcattc ctcccactca tgatctatag atctatagat
      541 ctctcgtggg atcattgttt ttctcttgat tcccactttg tggttctaag tactgtggtt
      601 tccaaatgtg tcagtttcat agcctgaaga acgagatcag cagcctctgt tccacataca
      661 cttcattctc agtattgttt tgccaagttc taattccatc agaagctggt cgagatccgg
      721 aacccttaat ataacttcgt ataatgtatg ctatacgtag ttattaggtc cctcgaagag
      781 gttcactagg cgcgcgaggc tcacggtcat gacagcaatt gcagaggaac ccagagtaat
      841 tgagactgac tgaccacctg acaagctgcc acggggaact gcagcttttg ctgaatagca
      901 ttttacagtg tttgttggaa acctgaattt ggttctgact tctgtggctg tttaaaatat
      961 agggcttgtg ggtcacttga aaagtacctg tagaagcccg gataacttga ggggaatgtc
     1021 tgtttgtact tttggaaatt atttaactgc ttggtttatc ctaggatcag aggctaaaca
     1081 actccatagt caacactttt ccacctgaca ttagaaccct gtaatgattt cattatggat
     1141 aggggaagtc taacaagaca atcgatttta aacagggttt taatcaaata agttcttccc
     1201 acttttaagc tgatgaaaga agtcattatt tcttgtgaat attttttcct ggagagcctt
     1261 atcatgtatt aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg
     1321 cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
     1381 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagtat
     1441 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1501 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1561 ttacatcgaa ctggatctca acagcggtaa gatccttgag agtttacgcc ccgaagaacg
     1621 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1681 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1741 ctcaccagtc acagaaaagc atctcacgga tggcatgaca gtaagagaat tatgcagtgc
     1801 tgccataacc atgagtgata acactgcggc caacttactt ctggcaacga tcggaggacc
     1861 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1921 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1981 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2041 acaattaata gactggatgg aggcggataa agttgcagga tcacttctgc gctcggccct
     2101 cccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2161 cattgcagca ctggggccag atggtaagcc ctcccgcatc gtagttatct acacgacggg
     2221 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2281 taagcattgg taatgagggc ccaaatgtaa tcacctggct caccttcggg tgggcctttc
     2341 tgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgatg
     2401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2821 gctgaagcca gttacctcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2941 caagaagatc ctttgatttt ctaccgaaga aaggcccacc cgtgaaggtg agccagtgag
     3001 ttgattgcag tccagttacg ctggagtctg aggctcgtcc tgaatgatat caagcttgaa
     3061 ttagttcctt tatttatctt taaacgccat ttaagttgat tacttatact caattatctt
     3121 actctaacaa attaggatcc ttcttaaata aagcaatgat ggcctttttt aaaacataga
     3181 ggtgtgaaag tttatagcat tttaaattta tttcaagagt agaagttact aaaatgattt
     3241 atgaaaataa tcttatttgg caatattacc ctggcttctg acatctaaga aaaggtggca
     3301 aatacaatat gctttactaa aaactagtag gtactcattt gcaaagacac taatgagtat
     3361 tggtgaatat gtgtaaccca tttatatctt gattacttgc agaactattt caagcctaga
     3421 agaaattgtt gaaaagcaag gagacaccat tgaatacctg aagcgacaca atgcgctgct
     3481 gagtaagcga ttgttggctc tcacttcctc agacctgggc tgtcagccaa gcaggacagg
     3541 aggctcagga ggctcaggag gctcatatcc ctatgacgtg cctgattacg ccggcacagg
     3601 atcctacccc tatgatgtgc ctgactacgc tggcagcgcc ggataccctt atgatgtgcc
     3661 tgattatgct ggagggagcg gctaagaatt ccgatcatat tcaataaccc ttaatataac
     3721 ttcgtataat gtatgctata cgaagttatt aggtctgaag aggagtttac gtccagccaa
     3781 gcttaggatc tcgacctcga aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     3841 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     3901 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     3961 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     4021 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     4081 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     4141 cgctttctgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
     4201 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     4261 cgacctgcat ccatctagat ctcgagcagc tgaagcttac catgaccgag tacaagccca
     4321 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4381 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4441 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4501 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4561 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4621 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4681 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgcc
//
