LOCUS       sequence-452493-        4366 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 4366)
  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 4366)
  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: pML499_SLC3A2-C-term_XTEN80-sfGFP-3xHA_pTMPLT
FEATURES             Location/Qualifiers
     source          1..4366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             789..812
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             828..1340
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVE"
     primer_bind     complement(867..888)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1475..1496
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1527..1607
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(2202..2218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2202..2218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2215..2237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2390..2978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2470..2489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3095..3119)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3219..4010,4011..4079))
                     /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     3842..3861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4080..4184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4283..4305
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4297..4314
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4298..4314
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggagcttact ggtgtcactt cggctgtaag tgccaacatg actgtgaagg taagagttct
       61 agatgggtag aaactgaccg gtggagggtg ggctgggctt gtagaagtct gtacccaggg
      121 gagcccctca cacgcttctg ctatgttttt atcattctta gggccagagt gaagaccctg
      181 gctccctcct ttccttgttc cggcggctga gtgaccagcg gagtaaggag cgctccctac
      241 tgcatgggga cttccacgcg ttctccgctg ggcctggact cttctcctat atccgccact
      301 gggaccagaa tgagcgtttt ctggtagtgc ttaactttgg ggatgtgggc ctctcggctg
      361 gactgcaggc ctccgacctg cctgccagcg ccagcctgcc agccaaggct gacctcctgc
      421 tcagcaccca gccaggccgt gaggagggct cccctcttga gctggaacgc ctgaaactgg
      481 agcctcacga agggctgctg ctccgcttcc cctacgcggc ctctggtggt acttccgggg
      541 gagggccgag ctctggcgca cccccaccaa gtggagggtc acctgccggg tccccaacat
      601 ctactgaaga aggcaccagc gaatccgcaa cgcccgagtc aggccctggt acctccacag
      661 aaccatctga aggtagtgcg cctggttccc cagctggaag ccctacttcc accgaagaag
      721 gcacgtcaac cgaaccaagt gaaggatctg cccctgggac cagcactgaa ccatctgagg
      781 gtggcggatt ggaagttttg tttcaaggtc caggatctaa cggcagcagc aagggcgagg
      841 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      901 agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt
      961 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1021 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcgccacgac ttcttcaagt
     1081 ccgccatgcc cgaaggctac gtccaggagc gcaccatcag cttcaaggac gacggcacct
     1141 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1201 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca
     1261 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1321 agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1381 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1441 tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1501 ccgccgggat cactgctgga gctggctatc catacgatgt tccagattac gcttatccgt
     1561 atgacgtccc ggactatgca tacccctatg atgtacccga ctacgccggt gcagggagtg
     1621 gaagttgact tcagcctgac atggacccac tacccttctc ctttccttcc caggcccttt
     1681 ggcttctgat ttttctcttt tttaaaaaca aacaaacaaa ctgttgcaga ttatgagtga
     1741 acccccaaat agggtgtttt ctgccttcaa ataaaagtca cccctgcatg gtgaagtctt
     1801 ccctctgctt ctctcatagc aggaatttct ctccttcctc ccctcccatg cgtctcctcc
     1861 aactccttca ttatttatgt agggtcccag ggccagagga gagggggcct cccaaagtca
     1921 acagcatgga ggtgtcaggg ctgggaaagc acccatgcct cacccatgct gggccacggg
     1981 gctctctgga catcttgggt cttgccgact cttaacttca tccccttcag caaagggctc
     2041 tagccaaaca tagttagttt tcctgttttt gcagacttgg cctttcacgg agagtagggg
     2101 tgggatggtc tggtttttaa aaggtcgtca gaagaccgtc agtgaaggct aggtggaggc
     2161 tcagtgatga taagtctgcg atggtggatg catgtgtcat ggtcatagct gtttcctgtg
     2221 tgaaattgtt atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag
     2281 acattaggtg gagttcagtt cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa
     2341 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2401 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2461 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2521 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2581 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2641 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2701 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2761 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2821 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2881 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2941 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3001 cggggtctga cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag
     3061 ggggcttcaa atcgtcctcg tgataccaat tcggagcctg cttttttgta caaacttgtt
     3121 gataatggca attcaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3361 gagccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4201 gtgccagata cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat
     4261 ccaccacaag cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtcatgca
     4321 taatccgcac gcatctggaa taaggaagtg ccattccgcc tgacct
//
