LOCUS       sequence-437950-        6922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Calreticulin truncated C 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 6922)
  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 6922)
  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 C
FEATURES             Location/Qualifiers
     source          1..6922
                     /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(2933..2951)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2934..2951)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2971..2988)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2977..3201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3247..3675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3334..3353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3544..3565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3684..3704)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3728..3938
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3750..3946
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3797..3932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3859..3878
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4013..4807
                     /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(4067..4086)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4677..4696
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4981..5102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5018..5037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5072..5091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5151..5167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5151..5167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5164..5186)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5175..5191
                     /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(5199..5205,5206..5223,5224..5229))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5244..5265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5382..5399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5553..6141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5633..5652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6312..6922,1..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 gcttaccgat aactttggcg tgctgggcct ggacctctgg
     1321 caggtcaagt ctggcaccat ctttgacaac ttcctcatca ccaacgatga ggcatacgct
     1381 gaggagtttg gcaacgagac gtggggcgta acaaaggcag cagagaaaca aatgaaggac
     1441 aaacaggacg aggagcagag gcttaaggag gaggaagaag acaagaaacg caaagaggag
     1501 gaggaggcag aggacaagga ggatgatgag gacaaagatg aggatgagga ggatgaggag
     1561 gacaaggagg aagatgagga ggaagatgtc cccggccagg ccagatctgc ggatcccgag
     1621 ggtgagtact tagcttcagc gctcctgcct ggacgcatcc cggctatgca gccccagtcc
     1681 agggcagcaa ggcaggcccc gtctgcctct tcacccggag gcctctgccc gccccactca
     1741 tgctcaggga gagggtcttc tggctttttc cccaggctct gggcaggcac aggctaggtg
     1801 cccctaaccc aggccctgca cacaaagggg caggtgctgg gctcagacct gccaagagcc
     1861 atatccggga ggaccctgcc cctgacctaa gcccacccca aaggccaaac tctccactcc
     1921 ctcagctcgg acaccttctc tcctcccaga ttccagtaac tcccaatctt ctctctgcag
     1981 agcccaaatc ttgtgacaaa actcacacat gcccaccgtg cccaggtaag ccagcccagg
     2041 cctcgccctc cagctcaagg cgggacaggt gccctagagt agcctgcatc cagggacagg
     2101 ccccagccgg gtgctgacac gtccacctcc atctcttcct cagcacctga actcctgggg
     2161 ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc
     2221 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac
     2281 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac
     2341 aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc
     2401 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc
     2461 tccaaagcca aaggtgggac ccgtggggtg cgagggccac atggacagag gccggctcgg
     2521 cccaccctct gccctgagag tgaccgctgt accaacctct gtccctacag ggcagccccg
     2581 agaaccacag gtgtacaccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag
     2641 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     2701 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     2761 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     2821 atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc
     2881 tccgggtaaa tgagtgcgac ggccgctcga gcatgcatct agagggccct attctatagt
     2941 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     3001 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3061 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3121 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3181 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3241 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3301 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3361 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3421 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3481 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3541 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3601 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3661 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3721 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3781 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3841 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3901 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3961 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4021 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4081 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4141 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4201 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4261 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4321 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4381 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4441 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4501 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4561 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4621 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4681 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4741 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4801 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4861 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4921 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5101 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5161 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5221 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5281 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6421 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6901 cgaccgagtt gctcttgccc gg
//
