LOCUS       sequence-437954-        6940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Calreticulin truncated P domain with C-terminal human 
            IgG1Fc tag in mammalian cell..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6940)
  AUTHORS   Sen Santara S, Lee DJ, Crespo A, Hu JJ, Walker C, Ma X, Zhang Y, 
            Chowdhury S, Meza-Sosa KF, Lewandrowski M, Zhang H, Rowe M, 
            McClelland A, Wu H, Junqueira C, Lieberman J
  TITLE     The NK cell receptor NKp46 recognizes ecto-calreticulin on 
            ER-stressed cells.
  JOURNAL   Nature. 2023 Apr;616(7956):348-356. doi: 10.1038/s41586-023-05912-0.
            Epub 2023 Apr 5.
  PUBMED    37020026
REFERENCE   2  (bases 1 to 6940)
  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"; doi: 
            "10.1038/s41586-023-05912-0"; journalName: "Nature"; date: 
            "2023-04"; volume: "616"; issue: "7956"; pages: "348-356"
COMMENT     Sequence Label: pCMV-CRT-Fc P
FEATURES             Location/Qualifiers
     source          1..6940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..32
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(251..355)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(434..453)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        625..1004
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1005..1208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1159..1179
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1253..1272
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1253..1271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2951..2969)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2952..2969)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2989..3006)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2995..3219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3265..3693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3352..3371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3562..3583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3702..3722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3746..3956
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3768..3964
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3815..3950
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3877..3896
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4031..4825
                     /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(4085..4104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4695..4714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4999..5120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5036..5055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5090..5109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5169..5185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5169..5185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5182..5204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5193..5209
                     /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(5217..5223,5224..5241,5242..5247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5262..5283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5400..5417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5571..6159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5651..5670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6330..6940,1..181,182..250))
                     /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"
ORIGIN
        1 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
       61 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      121 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      181 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      241 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      301 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      361 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctat
      421 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
      481 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
      541 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      601 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      661 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      721 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      781 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      841 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      901 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      961 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     1021 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     1081 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1141 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1201 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
     1261 ctcactatag ggagacccaa gcttaccgac gattgggact tcctgccacc caagaagata
     1321 aaggatcctg atgcttcaaa accggaagac tgggatgagc gggccaagat cgatgatccc
     1381 acagactcca agcctgagga ctgggacaag cccgagcata tccctgaccc tgatgctaag
     1441 aagcccgagg actgggatga agagatggac ggagagtggg aacccccagt gattcagaac
     1501 cctgagtaca agggtgagtg gaagccccgg cagatcgaca acccagatta caagggcact
     1561 tggatccacc cagaaattga caaccccgag tattctcccg atcccagtat ctatgcctat
     1621 agatctgcgg atcccgaggg tgagtactta gcttcagcgc tcctgcctgg acgcatcccg
     1681 gctatgcagc cccagtccag ggcagcaagg caggccccgt ctgcctcttc acccggaggc
     1741 ctctgcccgc cccactcatg ctcagggaga gggtcttctg gctttttccc caggctctgg
     1801 gcaggcacag gctaggtgcc cctaacccag gccctgcaca caaaggggca ggtgctgggc
     1861 tcagacctgc caagagccat atccgggagg accctgcccc tgacctaagc ccaccccaaa
     1921 ggccaaactc tccactccct cagctcggac accttctctc ctcccagatt ccagtaactc
     1981 ccaatcttct ctctgcagag cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc
     2041 caggtaagcc agcccaggcc tcgccctcca gctcaaggcg ggacaggtgc cctagagtag
     2101 cctgcatcca gggacaggcc ccagccgggt gctgacacgt ccacctccat ctcttcctca
     2161 gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc
     2221 ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac
     2281 cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag
     2341 ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac
     2401 caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc
     2461 cccatcgaga aaaccatctc caaagccaaa ggtgggaccc gtggggtgcg agggccacat
     2521 ggacagaggc cggctcggcc caccctctgc cctgagagtg accgctgtac caacctctgt
     2581 ccctacaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggatgagct
     2641 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     2701 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     2761 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     2821 gcaggggaac gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca
     2881 gaagagcctc tccctgtctc cgggtaaatg agtgcgacgg ccgctcgagc atgcatctag
     2941 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     3001 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3061 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3121 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3181 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3241 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3301 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3361 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3421 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3481 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3541 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3601 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3661 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3781 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3841 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3901 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3961 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4021 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4081 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4141 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4201 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4261 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4321 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4381 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4441 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4501 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4561 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4621 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4681 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4741 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4801 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4861 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4921 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4981 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5041 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5101 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5161 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5221 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
//
