LOCUS       sequence-437957-        7846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Calreticulin (full-length) 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 7846)
  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 7846)
  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 FL
FEATURES             Location/Qualifiers
     source          1..7846
                     /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(3857..3875)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3858..3875)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3895..3912)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3901..4125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4171..4599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4258..4277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4468..4489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4608..4628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4652..4862
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4674..4870
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4721..4856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4783..4802
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4937..5731
                     /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(4991..5010)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5601..5620
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5905..6026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5942..5961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5996..6015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6075..6091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6075..6091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6088..6110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6099..6115
                     /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(6123..6129,6130..6147,6148..6153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6168..6189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6306..6323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6477..7065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6557..6576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7236..7846,1..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 gcttaccatg ctgctatccg tgccgctgct gctcggcctc
     1321 ctcggcctgg ccgtcgccga gcctgccgtc tacttcaagg agcagtttct ggacggagac
     1381 gggtggactt cccgctggat cgaatccaaa cacaagtcag attttggcaa attcgttctc
     1441 agttccggca agttctacgg tgacgaggag aaagataaag gtttgcagac aagccaggat
     1501 gcacgctttt atgctctgtc ggccagtttc gagcctttca gcaacaaagg ccagacgctg
     1561 gtggtgcagt tcacggtgaa acatgagcag aacatcgact gtgggggcgg ctatgtgaag
     1621 ctgtttccta atagtttgga ccagacagac atgcacggag actcagaata caacatcatg
     1681 tttggtcccg acatctgtgg ccctggcacc aagaaggttc atgtcatctt caactacaag
     1741 ggcaagaacg tgctgatcaa caaggacatc cgttgcaagg atgatgagtt tacacacctg
     1801 tacacactga ttgtgcggcc agacaacacc tatgaggtga agattgacaa cagccaggtg
     1861 gagtccggct ccttggaaga cgattgggac ttcctgccac ccaagaagat aaaggatcct
     1921 gatgcttcaa aaccggaaga ctgggatgag cgggccaaga tcgatgatcc cacagactcc
     1981 aagcctgagg actgggacaa gcccgagcat atccctgacc ctgatgctaa gaagcccgag
     2041 gactgggatg aagagatgga cggagagtgg gaacccccag tgattcagaa ccctgagtac
     2101 aagggtgagt ggaagccccg gcagatcgac aacccagatt acaagggcac ttggatccac
     2161 ccagaaattg acaaccccga gtattctccc gatcccagta tctatgccta tgataacttt
     2221 ggcgtgctgg gcctggacct ctggcaggtc aagtctggca ccatctttga caacttcctc
     2281 atcaccaacg atgaggcata cgctgaggag tttggcaacg agacgtgggg cgtaacaaag
     2341 gcagcagaga aacaaatgaa ggacaaacag gacgaggagc agaggcttaa ggaggaggaa
     2401 gaagacaaga aacgcaaaga ggaggaggag gcagaggaca aggaggatga tgaggacaaa
     2461 gatgaggatg aggaggatga ggaggacaag gaggaagatg aggaggaaga tgtccccggc
     2521 caggccagat ctgcggatcc cgagggtgag tacttagctt cagcgctcct gcctggacgc
     2581 atcccggcta tgcagcccca gtccagggca gcaaggcagg ccccgtctgc ctcttcaccc
     2641 ggaggcctct gcccgcccca ctcatgctca gggagagggt cttctggctt tttccccagg
     2701 ctctgggcag gcacaggcta ggtgccccta acccaggccc tgcacacaaa ggggcaggtg
     2761 ctgggctcag acctgccaag agccatatcc gggaggaccc tgcccctgac ctaagcccac
     2821 cccaaaggcc aaactctcca ctccctcagc tcggacacct tctctcctcc cagattccag
     2881 taactcccaa tcttctctct gcagagccca aatcttgtga caaaactcac acatgcccac
     2941 cgtgcccagg taagccagcc caggcctcgc cctccagctc aaggcgggac aggtgcccta
     3001 gagtagcctg catccaggga caggccccag ccgggtgctg acacgtccac ctccatctct
     3061 tcctcagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc aaaacccaag
     3121 gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga cgtgagccac
     3181 gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca taatgccaag
     3241 acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt cctcaccgtc
     3301 ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa caaagccctc
     3361 ccagccccca tcgagaaaac catctccaaa gccaaaggtg ggacccgtgg ggtgcgaggg
     3421 ccacatggac agaggccggc tcggcccacc ctctgccctg agagtgaccg ctgtaccaac
     3481 ctctgtccct acagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga
     3541 tgagctgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga
     3601 catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc
     3661 cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag
     3721 gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta
     3781 cacgcagaag agcctctccc tgtctccggg taaatgagtg cgacggccgc tcgagcatgc
     3841 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     3901 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3961 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4021 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4081 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4141 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4201 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4261 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4321 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4381 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4441 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4501 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4561 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4621 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4681 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4801 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4861 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4921 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4981 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5041 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5101 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5161 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5221 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5281 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5341 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5401 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5461 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5521 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5581 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5641 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5701 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5761 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5821 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5881 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5941 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6001 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6061 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6121 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6181 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6241 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6301 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6361 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccgg
//
