LOCUS       sequence-390368-        6060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of ER-targeted metastable Halotag/folding 
            sensor (ER HaloM21K_F86L).
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_M21K_F86L_KDEL_pBABEpu
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..45
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             49..66
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1029..1049)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1034..1363
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1214..1349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1373..1972
                     /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(1373..1392)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1869..1889
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2193..2662
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2792..2809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2963..3551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3043..3062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3711..4502,4503..4571))
                     /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     4334..4353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4572..4676)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4707..5176
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5241..5440
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5641..6057
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5987..6009
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6025..6041
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 atgtctgcac ttctgatcct agctcttgtt ggagctgcag ttgctatgca ccatcatcat
       61 caccatgcgg ccgcatctct cgaggcagaa atcggtactg gctttccatt cgacccccat
      121 tatgtggaag tcctgggcga gcgcaagcac tacgtcgatg ttggtccgcg cgatggcacc
      181 cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg
      241 catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac
      301 aaaccagacc tgggttattt cttcgacgac cacgtccgct taatggatgc cttcatcgaa
      361 gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc
      421 cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc
      481 cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc
      541 accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg
      601 ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc
      661 ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt
      721 gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct
      781 gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct
      841 cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg
      901 ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct ttcgacggtc
      961 gacggatccg cagatctagc ggccaaggat gagttgtagg aattcgccag cacagtggtc
     1021 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     1081 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     1141 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     1201 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     1261 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     1321 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     1381 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     1441 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     1501 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     1561 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     1621 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     1681 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     1741 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     1801 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     1861 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     1921 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     1981 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     2041 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     2101 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     2161 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     2221 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     2281 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     2341 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     2401 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     2461 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     2521 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     2581 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     2641 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     2701 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     2761 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3661 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3721 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3781 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3841 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3901 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3961 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4021 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4081 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4141 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4201 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4261 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4321 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4381 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4441 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4501 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4561 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4681 tttccccgaa aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag
     4741 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     4801 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     4861 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     4921 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     4981 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     5041 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5101 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     5161 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     5221 ccgacccacc accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta
     5281 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc
     5341 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     5401 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     5461 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     5521 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     5581 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     5641 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     5701 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     5761 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     5821 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     5881 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     5941 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     6001 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc
//
