LOCUS       sequence-456019-        5382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology repair template for genome engineering. Organelle 
            endogenous marker expressing GFP and epitope tags for mass 
            spectrometry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5382)
  AUTHORS   Hein MY, Peng D, Todorova V, McCarthy F, Kim K, Liu C, Savy L, 
            Januel C, Baltazar-Nunez R, Sekhar M, Vaid S, Bax S, Vangipuram M, 
            Burgess J, Njoya L, Wang E, Ivanov IE, Byrum JR, Pradeep S, Gonzalez
            CG, Aniseia Y, Creery JS, McMorrow AH, Sunshine S, Yeung-Levy S, 
            DeFelice BC, Mehta SB, Itzhak DN, Elias JE, Leonetti MD
  TITLE     Global organelle profiling reveals subcellular localization and 
            remodeling at proteome scale.
  JOURNAL   Cell. 2024 Dec 26:S0092-8674(24)01344-8. doi: 
            10.1016/j.cell.2024.11.028.
  PUBMED    39742809
REFERENCE   2  (bases 1 to 5382)
  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: "Cell. 2024 
            Dec 26:S0092-8674(24)01344-8. doi: 10.1016/j.cell.2024.11.028."
COMMENT     Sequence Label: pML359_CAPRIN1-N-term_XTEN80-mEGFP-3xHA_pTMPLT
FEATURES             Location/Qualifiers
     source          1..5382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             548..628
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             join(641..643,644..1354)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(683..704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(944..963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1291..1312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1364..1387
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2488..2504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2488..2504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2501..2523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2512..2528
                     /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)."
     promoter        complement(join(2536..2542,2543..2560,2561..2566))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2581..2602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2719..2736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2890..3478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2970..2989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3649..4440,4441..4509))
                     /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     4272..4291
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4510..4614)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4682..4700
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4738..4760)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4860..4879
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5073..5095
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5087..5104
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5088..5104
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgcaggggaa gagggccgct gcgcccgggt gtcctttatc tccggacccg ctggagcttc
       61 cgaggggctt gcccccaagt ccccccaccc caccccggtc atttcccatg cccttctctg
      121 cccccaagcc ccgacctctc tctttcctcc cttctccgga agccggcaaa agccggccct
      181 ccccggtagg gaccgttgca gtggccgttg gcggccggcg cggcgatgcc ccgcctccgg
      241 cgcgagcctc ggagcccccg ggctggtagg ccgagccggc ggctccgcgg gggaggggcg
      301 ggtcgggggg cggtgcgagg cccagccggg cgccccgctg gcgggtcgcg cgcgcgcccg
      361 cgcactcttc ctgccctcga ggcgcgccgc gccccctccc ccacccgctc gcgtaggcta
      421 ccgctcgctg cgcgttttgt cccgcgtctc gccccgtccg tctcctgact ggccgctctt
      481 gtccttcctc ccgctttttc ttctctctcc ttgcggtctg aagatgccca gtggtggtac
      541 ttccgggtac ccatacgatg ttccagatta cgcttatccg tatgacgtcc cggactatgc
      601 atacccctac gatgtacccg attacgccgc tggagctggc gtgagcaagg gcgaggagct
      661 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      721 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      781 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
      841 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      901 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      961 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1021 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1081 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1141 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1201 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     1261 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1321 cgggatcact ctcggcatgg acgagctgta caagggtggc ggattggaag ttttgtttca
     1381 aggtccagga tctaacggca gcggagggcc gagctctggc gcacccccac caagtggagg
     1441 gtcacctgcc gggtccccaa catctactga agaaggcacc agcgaatccg caacgcccga
     1501 gtcaggccct ggtacctcca cagaaccatc tgaaggtagt gcgcctggtt ccccagctgg
     1561 aagccctact tccaccgaag aaggcacgtc aaccgaacca agtgaaggat ctgcccctgg
     1621 gaccagcact gaaccatctg agggtgcagg gagtggaagt tcggccacca gccacagcgg
     1681 gagcggcagc aagtcgtccg gaccgccacc gccgtcgggt tcctccggga gtgaggcggc
     1741 cgcgggagcc ggggccgccg cgccggcttc tcagcacccc gcaaccggca ccggcgctgt
     1801 ccagaccgag gccatgaagc agattctcgg ggtgatcgac aagaaacttc ggaacctgga
     1861 gaagaaaaag gtgccaggag ttggcgggga agggaggggt ggctgcggcc gggctctgcg
     1921 gcccctccga ccctggtcgc tggagccttc gcttcttttc gtctcgggag cgttactagg
     1981 tcccctcctc ccaccccctg gcccacacgc cccgtctcca cgttcagcgg gagcttcgtg
     2041 cccagaaaac gggctcttgg aagaggcact tgtcacctga ctcggacgcg gcactgtacc
     2101 ccaggcctct ttattactct tccgctgtgg gtccggcttt ttggccttta ggagtgggac
     2161 atgtgagggt gggggcctgt cgtcaggact tcactgtatg gtgcccttct ttccgtctct
     2221 gaggcccgat ttctctgacg agggcccaaa atgaagaggt ccctgcgcgg ggggctccgg
     2281 gttcaaccga ccgcctcgtg gagttggggc ggcctgcgtc ctgcagcctt ggggtctgtc
     2341 cgctcggtta ccatgcactc gagtcctgtc gagcgtcccc tcttcttccg taggagagaa
     2401 gtgtgtttag cgtcagaaga ccgtcagtga atcggatccc gggcccgtcg actgcagagg
     2461 cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2521 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2581 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2641 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2701 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2761 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2821 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2881 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2941 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3001 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3061 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3121 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3181 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3241 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3301 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3361 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3421 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3481 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3541 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3601 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3661 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3721 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3781 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3841 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3901 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3961 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4021 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4081 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4141 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4201 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4261 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4321 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4381 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4441 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4501 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4561 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4621 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4681 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4861 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     4921 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     4981 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5041 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     5101 cagtgaattc gagctcggta cctcgcgaat gcatctagat ggagcttact ggtgtcactt
     5161 cgcgacccag agggctgctg gctggctaag tccctcccgc tcccggctct cgcctcacta
     5221 ggagcggctc tcggtgcagc gggacagggc gaagcggcct gcgcccacgg agcgcgcgac
     5281 actgcccgga agggaccacc acccttgccc cctcagctgc ccactcgtga tttccagcgg
     5341 cctccgcgcg cgcacggtga gcgccacccg ggaccgagcc gc
//
