LOCUS       sequence-437956-        7309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human NKp46 (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 7309)
  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 7309)
  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-NKp46-Fc
FEATURES             Location/Qualifiers
     source          1..7309
                     /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(3320..3338)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3321..3338)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3358..3375)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3364..3588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3634..4062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3721..3740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3931..3952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4071..4091)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4115..4325
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4137..4333
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4184..4319
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4246..4265
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4400..5194
                     /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(4454..4473)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5064..5083
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5368..5489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5405..5424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5459..5478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5538..5554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5538..5554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5551..5573)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5562..5578
                     /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(5586..5592,5593..5610,5611..5616))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5631..5652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5769..5786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5940..6528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6020..6039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6699..7309,1..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 cagcagcaga ctctcccaaa accgttcatc
     1321 tgggccgagc cccatttcat ggttccaaag gaaaagcaag tgaccatctg ttgccaggga
     1381 aattatgggg ctgttgaata ccagctgcac tttgaaggaa gcctttttgc cgtggacaga
     1441 ccaaaacccc ctgagcggat taacaaagtc aaattctaca tcccggacat gaactcccgc
     1501 atggcagggc aatacagctg catctatcgg gttggggagc tctggtcaga gcccagcaac
     1561 ttgctggatc tggtggtaac agaaatgtat gacacaccca ccctctcggt tcatcctgga
     1621 cccgaagtga tctcgggaga gaaggtgacc ttctactgcc gtctagacac tgcaacaagc
     1681 atgttcttac tgctcaagga gggaagatcc agccacgtac agcgcggata cgggaaggtc
     1741 caggcggagt tccccctggg ccctgtgacc acagcccacc gagggacata ccgatgtttt
     1801 ggctcctata acaaccatgc ctggtctttc cccagtgagc cagtgaagct cctggtcaca
     1861 ggcgacattg agaacaccag ccttgcacct gaagacccca cctttcctga cacttggggc
     1921 acctaccttt taaccacaga gacgggactc cagaaagacc atgccctctg ggatcacact
     1981 gcccagaata gatctgcgga tcccgagggt gagtacttag cttcagcgct cctgcctgga
     2041 cgcatcccgg ctatgcagcc ccagtccagg gcagcaaggc aggccccgtc tgcctcttca
     2101 cccggaggcc tctgcccgcc ccactcatgc tcagggagag ggtcttctgg ctttttcccc
     2161 aggctctggg caggcacagg ctaggtgccc ctaacccagg ccctgcacac aaaggggcag
     2221 gtgctgggct cagacctgcc aagagccata tccgggagga ccctgcccct gacctaagcc
     2281 caccccaaag gccaaactct ccactccctc agctcggaca ccttctctcc tcccagattc
     2341 cagtaactcc caatcttctc tctgcagagc ccaaatcttg tgacaaaact cacacatgcc
     2401 caccgtgccc aggtaagcca gcccaggcct cgccctccag ctcaaggcgg gacaggtgcc
     2461 ctagagtagc ctgcatccag ggacaggccc cagccgggtg ctgacacgtc cacctccatc
     2521 tcttcctcag cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc
     2581 aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc
     2641 cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc
     2701 aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc
     2761 gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc
     2821 ctcccagccc ccatcgagaa aaccatctcc aaagccaaag gtgggacccg tggggtgcga
     2881 gggccacatg gacagaggcc ggctcggccc accctctgcc ctgagagtga ccgctgtacc
     2941 aacctctgtc cctacagggc agccccgaga accacaggtg tacaccctgc ccccatcccg
     3001 ggatgagctg accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag
     3061 cgacatcgcc gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc
     3121 tcccgtgctg gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag
     3181 caggtggcag caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca
     3241 ctacacgcag aagagcctct ccctgtctcc gggtaaatga gtgcgacggc cgctcgagca
     3301 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     3361 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3421 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3481 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3541 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3601 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3661 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3721 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3781 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3841 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3901 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3961 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4021 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4081 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4141 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4201 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4261 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4321 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4381 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4441 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4501 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4561 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4621 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4681 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4741 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4801 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4861 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4921 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4981 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5041 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5101 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5161 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5221 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5281 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5341 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5401 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5461 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5521 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5581 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5641 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5701 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5761 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5821 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6361 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6661 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6721 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6781 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6841 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6901 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6961 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7021 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7081 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7141 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7201 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7261 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccgg
//
