LOCUS       sequence-429203-        7787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human CALR ins5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7787)
  AUTHORS   De Marchi F, Okuda M, Morishita S, Imai M, Baba T, Horino M, Mori Y,
            Furuya C, Ogata S, Yang Y, Ando J, Ando M, Araki M, Komatsu N
  TITLE     Clinical and biological relevance of CREB3L1 in Philadelphia 
            chromosome-negative myeloproliferative neoplasms.
  JOURNAL   Leuk Res. 2022 Aug;119:106883. doi: 10.1016/j.leukres.2022.106883. 
            Epub 2022 May 31.
  PUBMED    35689957
REFERENCE   2  (bases 1 to 7787)
  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: "Leuk Res.";
            date: "2022-08"; volume: "119:106883. doi"; pages: " 
            10.1016/j.leukres.2022.106883. Epub 2022 May 31"
COMMENT     Sequence Label: pMSCV-IRES-mOrange/CALR ins5
FEATURES             Location/Qualifiers
     source          1..7787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    169..510
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             577..993
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     923..945
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     961..977
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2343..2892
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2487..2504)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2714..2733
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2899..3609
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     LTR             3770..4284
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4446..4462
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4470..4476,4477..4494,4495..4500))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4515..4536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4653..4670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4904..4923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5583..6374,6375..6443))
                     /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     6206..6225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6444..6548)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6616..6634
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6672..6694)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6794..6813
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7007..7029
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7157..7176
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7376..7787,1..105)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
       61 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
      121 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
      181 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
      241 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      301 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
      361 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
      421 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
      481 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
      541 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
      601 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
      661 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
      721 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
      781 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
      841 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
      901 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
      961 ctttatccag ccctcactcc ttctctaggc gccggaattc gccaccatgc tgctatccgt
     1021 gccgctgctg ctcggcctcc tcggcctggc cgtcgccgag cctgccgtct acttcaagga
     1081 gcagtttctg gacggagacg ggtggacttc ccgctggatc gaatccaaac acaagtcaga
     1141 ttttggcaaa ttcgttctca gttccggcaa gttctacggt gacgaggaga aagataaagg
     1201 tttgcagaca agccaggatg cacgctttta tgctctgtcg gccagtttcg agcctttcag
     1261 caacaaaggc cagacgctgg tggtgcagtt cacggtgaaa catgagcaga acatcgactg
     1321 tgggggcggc tatgtgaagc tgtttcctaa tagtttggac cagacagaca tgcacggaga
     1381 ctcagaatac aacatcatgt ttggtcccga catctgtggc cctggcacca agaaggttca
     1441 tgtcatcttc aactacaagg gcaagaacgt gctgatcaac aaggacatcc gttgcaagga
     1501 tgatgagttt acacacctgt acacactgat tgtgcggcca gacaacacct atgaggtgaa
     1561 gattgacaac agccaggtgg agtccggctc cttggaagac gattgggact tcctgccacc
     1621 caagaagata aaggatcctg atgcttcaaa accggaagac tgggatgagc gggccaagat
     1681 cgatgatccc acagactcca agcctgagga ctgggacaag cccgagcata tccctgaccc
     1741 tgatgctaag aagcccgagg actgggatga agagatggac ggagagtggg aacccccagt
     1801 gattcagaac cctgagtaca agggtgagtg gaagccccgg cagatcgaca acccagatta
     1861 caagggcact tggatccacc cagaaattga caaccccgag tattctcccg atcccagtat
     1921 ctatgcctat gataactttg gcgtgctggg cctggacctc tggcaggtca agtctggcac
     1981 catctttgac aacttcctca tcaccaacga tgaggcatac gctgaggagt ttggcaacga
     2041 gacgtggggc gtaacaaagg cagcagagaa acaaatgaag gacaaacagg acgaggagca
     2101 gaggcttaag gaggaggaag aagacaagaa acgcaaagag gaggaggagg cagaggacaa
     2161 ttgtcggagg atgatgagga caaagatgag gatgaggagg atgaggagga caaggaggaa
     2221 gatgaggagg aagatgtccc cggccaggcc aaggacgagc tgtagagagg cctgcctcca
     2281 gggctggact gaggcctgag aattcgttaa cctcgagcgg gatcaattcc gccccccccc
     2341 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2401 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2461 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2521 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2581 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2641 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2701 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2761 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     2821 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     2881 aacacgataa taccatgaat ggtgagcaag ggcgaggaga ataacatggc catcatcaag
     2941 gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga gttcgagatc
     3001 gagggcgagg gcgagggccg cccctacgag ggctttcaga ccgctaagct gaaggtgacc
     3061 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcatttcac ctacggctcc
     3121 aaggcctacg tgaagcaccc cgccgacatc cccgactact tcaagctgtc cttccccgag
     3181 ggcttcaagt gggagcgcgt gatgaactac gaggacggcg gcgtggtgac cgtgacccag
     3241 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     3301 ccctccgacg gccccgtgat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     3361 atgtaccccg aggacggtgc cctgaagggc aagatcaaga tgaggctgaa gctgaaggac
     3421 ggcggccact acacctccga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     3481 cccggcgcct acatcgtcga catcaagttg gacatcacct cccacaacga ggactacacc
     3541 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     3601 tacaagtaag cggccgcgac tctagagtcg acctgcaggc atgcaagctt caggtagccg
     3661 gctaacgtta acaaccggta cctctagaac tatagctagc atgcgcaaat ttaaagcgct
     3721 gatatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     3781 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     3841 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     3901 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     3961 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     4021 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     4081 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     4141 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg
     4201 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     4261 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct
     4321 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca
     4381 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg
     4441 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4501 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4561 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6601 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     6661 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     6721 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     6781 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     6841 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     6901 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6961 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7021 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     7081 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     7141 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     7201 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca
     7261 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag
     7321 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     7381 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     7441 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     7501 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     7561 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     7621 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     7681 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     7741 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgt
//
