LOCUS       sequence-429139-        6927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human CALR del52.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  TITLE     Masubuchi et al., 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6927)
  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            "Unpublished"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: pcDNA3.1/CALR del52
FEATURES             Location/Qualifiers
     source          1..6927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2518..2535)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2524..2748
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2794..3222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2881..2900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3091..3112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3231..3251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3236..3565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3416..3551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3478..3497
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3632..4426
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3686..3705)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4296..4315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4600..4721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4637..4656)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4691..4710
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4770..4786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4770..4786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4783..4805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4794..4810
                     /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(4818..4824,4825..4842,4843..4848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4863..4884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5001..5018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5172..5760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5252..5271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5931..6722,6723..6791))
                     /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     6554..6573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6792..6896)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgctgc tatccgtgcc gctgctgctc ggcctcctcg gcctggccgt cgccgagcct
     1021 gccgtctact tcaaggagca gtttctggac ggagacgggt ggacttcccg ctggatcgga
     1081 caaagatgag gatgaggagg atgaggagga caaggaggaa gatgaggagg aagatgtccc
     1141 cggccaggcc aaggacgagc tgtagagagg cctgcctcca gggctggact gaggcctgag
     1201 aattcgccac catgctgcta tccgtgccgc tgctgctcgg cctcctcggc ctggccgtcg
     1261 ccgagcctgc cgtctacttc aaggagcagt ttctggacgg agacgggtgg acttcccgct
     1321 ggatcgaatc caaacacaag tcagattttg gcaaattcgt tctcagttcc ggcaagttct
     1381 acggtgacga ggagaaagat aaaggtttgc agacaagcca ggatgcacgc ttttatgctc
     1441 tgtcggccag tttcgagcct ttcagcaaca aaggccagac gctggtggtg cagttcacgg
     1501 tgaaacatga gcagaacatc gactgtgggg gcggctatgt gaagctgttt cctaatagtt
     1561 tggaccagac agacatgcac ggagactcag aatacaacat catgtttggt cccgacatct
     1621 gtggccctgg caccaagaag gttcatgtca tcttcaacta caagggcaag aacgtgctga
     1681 tcaacaagga catccgttgc aaggatgatg agtttacaca cctgtacaca ctgattgtgc
     1741 ggccagacaa cacctatgag gtgaagattg acaacagcca ggtggagtcc ggctccttgg
     1801 aagacgattg ggacttcctg ccacccaaga agataaagga tcctgatgct tcaaaaccgg
     1861 aagactggga tgagcgggcc aagatcgatg atcccacaga ctccaagcct gaggactggg
     1921 acaagcccga gcatatccct gaccctgatg ctaagaagcc cgaggactgg gatgaagaga
     1981 tggacggaga gtgggaaccc ccagtgattc agaaccctga gtacaagggt gagtggaagc
     2041 cccggcagat cgacaaccca gattacaagg gcacttggat ccacccagaa attgacaacc
     2101 ccgagtattc tcccgatccc agtatctatg cctatgataa ctttggcgtg ctgggcctgg
     2161 acctctggca ggtcaagtct ggcaccatct ttgacaactt cctcatcacc aacgatgagg
     2221 catacgctga ggagtttggc aacgagacgt ggggcgtaac aaaggcagca gagaaacaaa
     2281 tgaaggacaa acaggacgag gagcagagga caaggaggat gatgaggaca aagatgagga
     2341 tgaggaggat gaggaggaca aggaggaaga tgaggaggaa gatgtccccg gccaggccaa
     2401 ggacgagctg tagagaggcc tgcctccagg gctggactga ggcctgagaa ttctgcagat
     2461 atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     2521 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2581 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2641 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2701 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2761 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2821 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2881 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2941 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3001 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3061 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3121 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3181 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3241 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3301 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3361 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3421 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3481 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3541 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3601 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3661 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3721 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3781 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3841 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3901 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3961 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4021 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4081 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4141 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4201 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4261 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4321 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4381 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4441 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4501 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4561 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4621 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4681 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4741 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4801 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4921 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4981 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5581 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5881 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5941 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6001 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6061 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6121 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6181 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6241 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6301 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6361 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6421 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6481 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6541 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6601 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6661 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6721 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6781 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6841 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6901 tttccccgaa aagtgccacc tgacgtc
//
