LOCUS       sequence-398281-        7033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ""soluble"" HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  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 7033)
  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: pJPT001-HaloTag-1x FLAG
FEATURES             Location/Qualifiers
     source          1..7033
                     /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             46..930
                     /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             940..963
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1016..1137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1027..1046)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1081..1100
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1252..1298
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1328..1464
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1509..1580
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1806..2394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1886..1905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2894..4103
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3940..3959
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4107..4706
                     /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(4107..4126)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4603..4623
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4733..4781
                     /note="synthetic polyadenylation signal"
     CDS             join(5047..5115,5116..5907)
                     /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(5265..5284)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5920..6056
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6208..6279
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6284..6330)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6348..6396
                     /note="synthetic polyadenylation signal"
     enhancer        6477..6780
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6782..6984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6935..6955
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg gaattcggcg gctccgaaat cggtactggc
       61 tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt
      121 ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg
      181 tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc
      241 ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc
      301 atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg
      361 ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca
      421 tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag
      481 accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt
      541 tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac
      601 cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac
      661 gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac
      721 tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc
      781 ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc
      841 ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg
      901 cgctggctgt ctactctgga gatttccggc tccggatccg actacaagga cgatgacgac
      961 aagtgagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1021 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1081 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1141 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1201 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1261 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1321 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1381 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     1441 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     1501 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     1561 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     1621 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     1681 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     1741 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     2641 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     2701 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     2761 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     2821 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     2881 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     2941 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     3001 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     3061 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     3121 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     3181 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     3241 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     3301 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     3361 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     3421 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     3481 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     3541 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     3601 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     3661 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     3721 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     3781 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     3841 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     3901 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     3961 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     4021 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     4081 cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg cgcctcgcca
     4141 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4201 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4261 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4321 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4381 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggagggcc
     4441 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     4501 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg
     4561 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     4621 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     4681 gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag ataataaaat
     4741 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     4801 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     4861 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     4921 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     4981 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     5041 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5101 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5161 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5221 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5281 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5341 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5401 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5461 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5521 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5581 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     5641 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     5701 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5761 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5821 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5881 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     5941 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     6001 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     6061 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     6121 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     6181 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     6241 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     6301 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     6361 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     6421 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6481 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6541 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6601 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6661 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6721 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     6781 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     6841 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     6901 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     6961 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
     7021 ctaccatcca ctc
//
