LOCUS       sequence-452503-        4893 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 4893)
  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 4893)
  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: pML340_RAB11A-N-term_XTEN80-mEGFP-3xHA_pTMPLT
FEATURES             Location/Qualifiers
     source          1..4893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..28
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(247..351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     450..472
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     464..481
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     465..481
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1323..1403
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             join(1416..1418,1419..2129)
                     /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(1458..1479)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1719..1738)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2066..2087
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2139..2162
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3262..3278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3262..3278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3275..3297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3450..4038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3530..3549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(4155..4179)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4279..4893,1..177,178..246))
                     /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"
ORIGIN
        1 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
       61 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
      121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
      181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
      241 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
      301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
      361 ccgaaaagtg ccagatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
      421 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      481 tcatgcataa tccgcacgca tctggaataa ggaagtgcca ttccgcctga cctggagctt
      541 actggtgtca cttcaaagcc cagggttaaa aaaaaggcag tgaagaagct cattaagaca
      601 accccccgcc aggtagcaga tagcacctga aatgcagtgg gaaatttgtg aggtcttcat
      661 aggcaaggat taaattttac atctccatct taagatgtgt catagttgtt caattactta
      721 gtgaccaaaa aagtgaatta aagatgttgt caagattaaa aaaacaaaaa ccaatgcagg
      781 ttccaccgag atttgaactc ggatcgctgg attcagagtc cagagtgcta accattacac
      841 catggaaccg ccacgcatgt gtagctgcct tcggctgtct aatcctcaga gaaccccgcc
      901 cccatccaca aacccaccac tcacaggcgg tcccgcctgg ttccagcgag ccgcttccgg
      961 cacggtagct cgagaaatga gcaagcggcc actaagacta tggtagctag gagttccagg
     1021 actcagtttc ccctttgagc ctcctttagc gactaaagct tgaagcccca cgcatctcga
     1081 ctctcgcgca caccgccctt gttgggctca ggggcggggc gccgcccccg gaagtacttc
     1141 cccttaaagg ctggggcctg ccggaaatgg cgcagcggca gggaggggct cttcacccag
     1201 tccggcagtt gaagctcggc gctcgggtta cccctgcagc gacgccccct ggtcccacag
     1261 ataccactgc tgctcccgcc ctttcgctcc tcggccgcgc aatgagtggt ggtacttccg
     1321 ggtacccata cgatgttcca gattacgctt atccgtatga cgtcccggac tatgcatacc
     1381 cctacgatgt acccgattac gccgctggag ctggcgtgag caagggcgag gagctgttca
     1441 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1501 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1561 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1621 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1681 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1741 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1801 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1861 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1921 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1981 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     2041 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2101 tcactctcgg catggacgag ctgtacaagg gtggcggatt ggaagttttg tttcaaggtc
     2161 caggatctaa cggcagcgga gggccgagct ctggcgcacc cccaccaagt ggagggtcac
     2221 ctgccgggtc cccaacatct actgaagaag gcaccagcga atccgcaacg cccgagtcag
     2281 gccctggtac ctccacagaa ccatctgaag gtagtgcgcc tggttcccca gctggaagcc
     2341 ctacttccac cgaagaaggc acgtcaaccg aaccaagtga aggatctgcc cctgggacca
     2401 gcactgaacc atctgagggt gcagggagtg gaagtggcac ccgcgacgac gagtacgact
     2461 acctctttaa aggtgaggcc atgggctctc gcactctaca cagtcctcgt tcggggaccc
     2521 gggccactcc cggtggaccc tcgtgccggc cacccctgca ctgatatagg cctccctcag
     2581 cccttccttt ttgtgcggtt ccgtctccta cccagctcag cctcttctcc cccgctcaga
     2641 caggggtccc catcacatgc cgctctctga gcgacctctc cataggcctt cgctggcctc
     2701 agagcccctc cctgcgtgtc cttcccctgg cggactgcct tctcccacat cgtcgaattc
     2761 ctttccccgg gttctacggc cccgccgctc ctcccaccat ctctcttttc gggtgtagcg
     2821 ccccctcccc ctcggcgtac acccttccca gctcgcgtcc tctcccgaag cccctctgac
     2881 gggttcttcg cttccctctt ggccttgcct tcggtgcaga ctcccattac aggtcttttt
     2941 cttatctgcc ctctttcccc cctcgagcga gcgtctcttc tgtatctccc ctccacttgg
     3001 acggccgccc ctctaagctt ctcccatccc gggtccagga ccccgtaaaa tctgagcggg
     3061 ccaagggcct ccgcccccac cctccgcccc gcggttctcg cggtccttaa ccttctctga
     3121 ccgctccctc tcccactccc catttggcaa ctctctctac cttccctccc caaatcgtgt
     3181 ttagacgtca gaagaccgtc agtgaaggct aggtggaggc tcagtgatga taagtctgcg
     3241 atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcag
     3301 agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg gagttcagtt
     3361 cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4021 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctctattc
     4081 aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa atcgtcctcg
     4141 tgataccaat tcggagcctg cttttttgta caaacttgtt gataatggca attcaaggat
     4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4381 gggcttacca tctggcccca gtgctgcaat gataccgcga gagccacgct caccggctcc
     4441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4861 tatgcggcga ccgagttgct cttgcccggc gtc
//
