LOCUS       sequence-429012-        7172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human CALR ins5-linker-Venus-KDEL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  TITLE     Masubuchi et al., 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7172)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMSCV/CALR ins5-linker-Venus-KDEL
FEATURES             Location/Qualifiers
     source          1..7172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    317..658
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             725..1141
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1071..1093
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1109..1125
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(2475..2477,2478..2480,2481..3191)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2520..2541)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2781..2800)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3128..3149
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3303..3817
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3979..3995
                     /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(4003..4009,4010..4027,4028..4033))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4048..4069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4186..4203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4357..4945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4437..4456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5116..5907,5908..5976))
                     /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     5739..5758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5977..6081)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6149..6167
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6205..6227)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6327..6346
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6540..6562
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6690..6709
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6909..7172,1..253)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
       61 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
      121 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
      181 gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact gcccacctcg
      241 ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg gaccaccgac
      301 ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg
      361 tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta actagctctg
      421 tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag
      481 acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg
      541 gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc
      601 gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca
      661 gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
      721 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
      781 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
      841 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
      901 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
      961 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1021 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1081 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1141 cggaattcgc caccatgctg ctatccgtgc cgctgctgct cggcctcctc ggcctggccg
     1201 tcgccgagcc tgccgtctac ttcaaggagc agtttctgga cggagacggg tggacttccc
     1261 gctggatcga atccaaacac aagtcagatt ttggcaaatt cgttctcagt tccggcaagt
     1321 tctacggtga cgaggagaaa gataaaggtt tgcagacaag ccaggatgca cgcttttatg
     1381 ctctgtcggc cagtttcgag cctttcagca acaaaggcca gacgctggtg gtgcagttca
     1441 cggtgaaaca tgagcagaac atcgactgtg ggggcggcta tgtgaagctg tttcctaata
     1501 gtttggacca gacagacatg cacggagact cagaatacaa catcatgttt ggtcccgaca
     1561 tctgtggccc tggcaccaag aaggttcatg tcatcttcaa ctacaagggc aagaacgtgc
     1621 tgatcaacaa ggacatccgt tgcaaggatg atgagtttac acacctgtac acactgattg
     1681 tgcggccaga caacacctat gaggtgaaga ttgacaacag ccaggtggag tccggctcct
     1741 tggaagacga ttgggacttc ctgccaccca agaagataaa ggatcctgat gcttcaaaac
     1801 cggaagactg ggatgagcgg gccaagatcg atgatcccac agactccaag cctgaggact
     1861 gggacaagcc cgagcatatc cctgaccctg atgctaagaa gcccgaggac tgggatgaag
     1921 agatggacgg agagtgggaa cccccagtga ttcagaaccc tgagtacaag ggtgagtgga
     1981 agccccggca gatcgacaac ccagattaca agggcacttg gatccaccca gaaattgaca
     2041 accccgagta ttctcccgat cccagtatct atgcctatga taactttggc gtgctgggcc
     2101 tggacctctg gcaggtcaag tctggcacca tctttgacaa cttcctcatc accaacgatg
     2161 aggcatacgc tgaggagttt ggcaacgaga cgtggggcgt aacaaaggca gcagagaaac
     2221 aaatgaagga caaacaggac gaggagcaga ggcttaagga ggaggaagaa gacaagaaac
     2281 gcaaagagga ggaggaggca gaggacaatt gtcggaggat gatgaggaca aagatgagga
     2341 tgaggaggat gaggaggaca aggaggaaga tgaggaggaa gatgtccccg gccaggccaa
     2401 ggacgagctg tagagaggcc tgcctccagg gctggactga ggccggtgga ggtggatctg
     2461 gtggtggagg atcaatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2521 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2581 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     2641 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     2701 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2761 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2821 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2881 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     2941 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     3001 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3061 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     3121 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3181 agctgtacaa gaaggacgag ctgtagaccg gtacctctag aactatagct agcatgcgca
     3241 aatttaaagc gctgatatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3301 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     3361 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     3421 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3481 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     3541 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3601 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3661 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     3721 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3781 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     3841 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     3901 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     3961 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4861 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4921 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4981 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5041 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5101 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5161 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5221 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5281 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5341 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5401 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5461 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5521 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5581 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5641 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5701 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5761 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5821 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5881 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5941 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6001 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6061 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6121 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6181 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6241 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6301 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6361 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6421 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6481 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6541 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     6601 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6661 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6721 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     6781 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     6841 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     6901 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     6961 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
     7021 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     7081 agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca
     7141 tcagatgttt ccagggtgcc ccaaggacct ga
//
