LOCUS       sequence-390365-        6060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of ER-targeted metastable Halotag/folding 
            sensor (HaloK73T).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6060)
  AUTHORS   Melo EP, Konno T, Farace I, Awadelkareem MA, Skov LR, Teodoro F, 
            Sancho TP, Paton AW, Paton JC, Fares M, Paulo PMR, Zhang X, Avezov E
  TITLE     Stress-induced protein disaggregation in the endoplasmic reticulum 
            catalysed by BiP.
  JOURNAL   Nat Commun. 2022 May 6;13(1):2501. doi: 10.1038/s41467-022-30238-2.
  PUBMED    35523806
REFERENCE   2  (bases 1 to 6060)
  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: "2022-05-6"; volume: "13(1):2501. doi"; pages: " 
            10.1038/s41467-022-30238-2"
COMMENT     Sequence Label: SP_H6_Halo_K73T_KDEL_pBABEpu
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    523..722
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             923..1339
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1269..1291
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1307..1323
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1343..1387
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1391..1408
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2371..2391)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2376..2705
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2556..2691
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2715..3314
                     /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(2715..2734)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3211..3231
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3535..4004
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4134..4151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4305..4893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4385..4404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5053..5844,5845..5913))
                     /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     5676..5695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5914..6018)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6049..6060,1..458)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
       61 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
      121 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
      181 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
      241 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
      301 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
      361 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
      421 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
      481 ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta
      541 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
      601 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      661 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
      721 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
      781 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
      841 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      901 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
      961 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1021 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1081 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1141 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1201 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1261 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1321 cactccttct ctaggcgccg gcatgtctgc acttctgatc ctagctcttg ttggagctgc
     1381 agttgctatg caccatcatc atcaccatgc ggccgcatct ctcgaggcag aaatcggtac
     1441 tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga
     1501 tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta
     1561 cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct
     1621 gatcggtatg ggcaaatccg acacaccaga cctgggttat ttcttcgacg accacgtccg
     1681 cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga
     1741 ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat
     1801 tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg
     1861 cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcagaa
     1921 cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat
     1981 ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc
     2041 aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat
     2101 ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct
     2161 gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga
     2221 catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat
     2281 cgcgcgctgg ctttcgacgg tcgacggatc cgcagatcta gcggccaagg atgagttgta
     2341 ggaattcgcc agcacagtgg tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2401 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2461 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2521 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2581 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2641 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2701 gcaaaaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     2761 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     2821 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     2881 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     2941 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     3001 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     3061 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     3121 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     3181 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     3241 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     3301 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     3361 cccatgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     3421 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     3481 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     3541 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3601 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3661 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     3721 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     3781 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     3841 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     3901 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     3961 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     4021 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     4081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4141 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4261 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4321 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4381 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4441 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4501 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4561 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4621 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4681 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4741 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4801 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4861 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4921 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4981 aggatcttca cctagatcct tttgcggccg caaatcaatc taaagtatat atgagtaaac
     5041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcagcct gaatatgggc
//
