LOCUS       sequence-398277-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ""PM"" HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  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 7132)
  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: pJPT003-PM-HaloTag-1x FLAG
FEATURES             Location/Qualifiers
     source          1..7132
                     /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             145..1029
                     /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             1039..1062
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1115..1236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1126..1145)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1180..1199
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1351..1397
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1427..1563
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1608..1679
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1905..2493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1985..2004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2993..4202
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4039..4058
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4206..4805
                     /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(4206..4225)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4702..4722
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4832..4880
                     /note="synthetic polyadenylation signal"
     CDS             join(5146..5214,5215..6006)
                     /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(5364..5383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6019..6155
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6307..6378
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6383..6429)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6447..6495
                     /note="synthetic polyadenylation signal"
     enhancer        6576..6879
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6881..7083
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7034..7054
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg ggctgcatca agagcaagcg gaaggacaag
       61 gacctggaac tgaagctgcg gatcctgcag agcaccgtgc ctagagctag agatcctcca
      121 gtggccacag aattcggcgg ctccgaaatc ggtactggct ttccattcga cccccattat
      181 gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga tggcacccct
      241 gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat catcccgcat
      301 gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa atccgacaaa
      361 ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt catcgaagcc
      421 ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct gggtttccac
      481 tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt catccgccct
      541 atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc cttccgcacc
      601 accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg tacgctgccg
      661 atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga gccgttcctg
      721 aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat cgccggtgag
      781 ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca gtcccctgtc
      841 ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga agccgctcgc
      901 ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct gaatctgctg
      961 caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc tactctggag
     1021 atttccggct ccggatccga ctacaaggac gatgacgaca agtgagctag cttgactgac
     1081 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
     1141 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1201 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1261 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1321 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     1381 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     1441 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     1501 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     1561 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     1621 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     1681 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
     1741 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
     1801 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
     1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2341 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2701 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     2761 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     2821 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     2881 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     2941 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg
     3001 cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca
     3061 gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     3121 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     3181 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     3241 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     3301 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
     3361 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
     3421 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     3481 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     3541 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     3601 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc
     3661 gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt
     3721 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     3781 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     3841 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     3901 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     3961 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     4021 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     4081 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     4141 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     4201 acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4261 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4321 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4381 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4441 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4501 gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg caccggccca
     4561 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4621 tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg gtgcccgcct
     4681 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4741 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4801 cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat
     4861 gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc
     4921 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     4981 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     5041 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     5101 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     5161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     5281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     5341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     5461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5521 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     5581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5701 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     5761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6001 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     6061 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     6121 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     6181 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     6241 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     6301 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     6361 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     6421 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
     6481 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
     6541 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     6601 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     6661 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     6721 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     6781 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     6841 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
     6901 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     6961 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     7021 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     7081 gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac tc
//
