LOCUS       sequence-437952-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Calreticulin truncated N 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 7146)
  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 7146)
  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 N
FEATURES             Location/Qualifiers
     source          1..7146
                     /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(3157..3175)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3158..3175)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3195..3212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3201..3425
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3471..3899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3768..3789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3908..3928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3952..4162
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3974..4170
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4021..4156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4083..4102
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4237..5031
                     /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(4291..4310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4901..4920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5205..5326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5242..5261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5296..5315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5375..5391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5375..5391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5388..5410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5399..5415
                     /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(5423..5429,5430..5447,5448..5453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5468..5489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5606..5623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5777..6365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5857..5876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6536..7146,1..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 gcttacgagc ctgccgtcta cttcaaggag cagtttctgg
     1321 acggagacgg gtggacttcc cgctggatcg aatccaaaca caagtcagat tttggcaaat
     1381 tcgttctcag ttccggcaag ttctacggtg acgaggagaa agataaaggt ttgcagacaa
     1441 gccaggatgc acgcttttat gctctgtcgg ccagtttcga gcctttcagc aacaaaggcc
     1501 agacgctggt ggtgcagttc acggtgaaac atgagcagaa catcgactgt gggggcggct
     1561 atgtgaagct gtttcctaat agtttggacc agacagacat gcacggagac tcagaataca
     1621 acatcatgtt tggtcccgac atctgtggcc ctggcaccaa gaaggttcat gtcatcttca
     1681 actacaaggg caagaacgtg ctgatcaaca aggacatccg ttgcaaggat gatgagttta
     1741 cacacctgta cacactgatt gtgcggccag acaacaccta tgaggtgaag attgacaaca
     1801 gccaggtgga gtccggctcc ttggaaagat ctgcggatcc cgagggtgag tacttagctt
     1861 cagcgctcct gcctggacgc atcccggcta tgcagcccca gtccagggca gcaaggcagg
     1921 ccccgtctgc ctcttcaccc ggaggcctct gcccgcccca ctcatgctca gggagagggt
     1981 cttctggctt tttccccagg ctctgggcag gcacaggcta ggtgccccta acccaggccc
     2041 tgcacacaaa ggggcaggtg ctgggctcag acctgccaag agccatatcc gggaggaccc
     2101 tgcccctgac ctaagcccac cccaaaggcc aaactctcca ctccctcagc tcggacacct
     2161 tctctcctcc cagattccag taactcccaa tcttctctct gcagagccca aatcttgtga
     2221 caaaactcac acatgcccac cgtgcccagg taagccagcc caggcctcgc cctccagctc
     2281 aaggcgggac aggtgcccta gagtagcctg catccaggga caggccccag ccgggtgctg
     2341 acacgtccac ctccatctct tcctcagcac ctgaactcct ggggggaccg tcagtcttcc
     2401 tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg
     2461 tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg
     2521 tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg
     2581 tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca
     2641 aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaaggtg
     2701 ggacccgtgg ggtgcgaggg ccacatggac agaggccggc tcggcccacc ctctgccctg
     2761 agagtgaccg ctgtaccaac ctctgtccct acagggcagc cccgagaacc acaggtgtac
     2821 accctgcccc catcccggga tgagctgacc aagaaccagg tcagcctgac ctgcctggtc
     2881 aaaggcttct atcccagcga catcgccgtg gagtgggaga gcaatgggca gccggagaac
     2941 aactacaaga ccacgcctcc cgtgctggac tccgacggct ccttcttcct ctacagcaag
     3001 ctcaccgtgg acaagagcag gtggcagcag gggaacgtct tctcatgctc cgtgatgcat
     3061 gaggctctgc acaaccacta cacgcagaag agcctctccc tgtctccggg taaatgagtg
     3121 cgacggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     3181 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3241 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3301 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3361 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3421 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3601 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3661 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3781 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3841 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3901 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4021 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4081 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4141 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4201 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4261 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4321 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4381 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4441 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4501 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4561 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4621 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4681 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4741 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4801 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4861 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4921 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4981 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5041 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5101 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5161 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5341 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5401 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5461 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5521 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5581 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5641 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5701 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5761 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5821 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5881 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5941 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6001 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6061 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6121 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6181 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6241 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6301 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6361 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6421 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6481 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6541 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6601 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6661 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6721 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6781 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6841 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6901 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6961 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7021 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7081 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7141 gcccgg
//
