LOCUS       sequence-398268-        7153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express dCore Lat mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  AUTHORS   Peruzzi JA, Gunnels TF, Edelstein HI, Lu P, Baker D, Leonard JN, 
            Kamat NP
  TITLE     Enhancing extracellular vesicle cargo loading and functional 
            delivery by engineering protein-lipid interactions.
  JOURNAL   Nat Commun. 2024 Jul 4;15(1):5618. doi: 10.1038/s41467-024-49678-z.
  PUBMED    38965227
REFERENCE   2  (bases 1 to 7153)
  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: "Nat 
            Commun."; date: "2024-07-4"; volume: "15(1):5618. doi"; pages: " 
            10.1038/s41467-024-49678-z"
COMMENT     Sequence Label: pJPT020-LAT_dCore-HaloTag-3x FLAG
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      22..31
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             124..1008
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             1018..1083
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1136..1257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1147..1166)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1201..1220
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1372..1418
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1448..1584
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1629..1700
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1926..2514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2006..2025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3014..4223
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4060..4079
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4227..4826
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4227..4246)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4723..4743
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4853..4901
                     /note="synthetic polyadenylation signal"
     CDS             join(5167..5235,5236..6027)
                     /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(5385..5404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6040..6176
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6328..6399
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6404..6450)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6468..6516
                     /note="synthetic polyadenylation signal"
     enhancer        6597..6900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6902..7104
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7055..7075
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg gaggaggcca tcctggtccc ctgcgtgctg
       61 gggctcgcca tgttgatggc actgtgtgtg cactgccaca gactgccagg atccggcggc
      121 tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc
      181 atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac
      241 ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc
      301 attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc
      361 gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc
      421 ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag
      481 cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg
      541 ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg
      601 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
      661 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
      721 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
      781 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
      841 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
      901 tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg
      961 atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggctc cgaattcgat
     1021 tacaaggacc acgatggcga ctataaggat cacgacatcg actacaagga cgatgacgac
     1081 aagtgagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1141 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1201 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1261 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1321 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1381 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1441 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1501 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     1561 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     1621 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     1681 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     1741 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     1801 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     1861 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2101 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2341 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     2761 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     2821 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     2881 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     2941 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3001 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     3061 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     3121 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     3181 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     3241 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     3301 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     3361 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     3421 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     3481 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     3541 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     3601 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     3661 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     3721 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     3781 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     3841 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     3901 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     3961 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     4021 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     4081 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     4141 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     4201 cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg cgcctcgcca
     4261 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4321 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4381 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4441 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4501 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggagggcc
     4561 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     4621 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg
     4681 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     4741 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     4801 gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag ataataaaat
     4861 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     4921 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     4981 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     5041 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     5101 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     5161 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5281 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5341 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5401 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5461 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5581 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5701 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     5761 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     5821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5881 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5941 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6001 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     6061 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     6121 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     6181 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     6241 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     6301 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     6361 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     6421 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     6481 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     6541 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6601 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6661 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6721 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6781 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6841 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     6901 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     6961 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     7021 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     7081 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
     7141 ctaccatcca ctc
//
