LOCUS       sequence-471033-        5784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminal His-tagged human SMSr (SAMD8) protein in insect 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  AUTHORS   Murakami C, Sakane F
  TITLE     Sphingomyelin synthase-related protein generates diacylglycerol via 
            the hydrolysis of glycerophospholipids in the absence of ceramide.
  JOURNAL   J Biol Chem. 2021 Jan-Jun;296:100454. doi: 
            10.1016/j.jbc.2021.100454. Epub 2021 Feb 20.
  PUBMED    33621517
REFERENCE   2  (bases 1 to 5784)
  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"; journalName: "J Biol 
            Chem. 2021 Jan-Jun;296:100454. doi: 10.1016/j.jbc.2021.100454. Epub 
            2021 Feb 20."
COMMENT     Sequence Label: pOET3-6xHis-hSMSr
FEATURES             Location/Qualifiers
     source          1..5784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_recomb     949..1776
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             949..1073
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(1111..1716)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     CDS             1884..1901
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1902..1916
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     3'UTR           3203..3575
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3927..3944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4037..4625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4117..4136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4796..5587,5588..5656))
                     /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"
     primer_bind     5419..5438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5657..5761)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccgtgac gttaaaacta ttaagccatc caatcgaccg ttagtcgaat caggaccgct
      421 ggtgcgagaa gccgcgaagt atggcgaatg catcgtataa cgtgtggagt ccgctcatta
      481 gagcgtcatg tttagacaag aaagctacat atttaattga tcccgatgat tttattgata
      541 aattgaccct aactccatac acggtattct acaatggcgg ggttttggtc aaaatttccg
      601 gactgcgatt gtacatgctg ttaacggctc cgcccactat taatgaaatt aaaaattcca
      661 attttaaaaa acgcagcaag agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa
      721 atgtcgtgga catgctgaac aacaagatta atatgcctcc gtgtataaaa aaaatattga
      781 acgatttgaa agaaaacaat gtaccgcgcg gcggtatgta caggaagagg tttatactaa
      841 actgttacat tgcaaacgtg gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg
      901 ctctgacgca tttctacaac cacgactcca agtgtgtggg tgaagtcatg catcttttaa
      961 tcaaatccca agatgtgtat aaaccaccaa actgccaaaa aatgaaaact gtggacaagc
     1021 tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat ttgtaattat tgaataataa
     1081 aacaattata aatgctaaat ttgtttttta ttaacgatac aaaccaaacg caacaagaac
     1141 atttgtagta ttatctataa ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa
     1201 atcattttca aatgattcac agttaatttg cgacaatata attttatttt cacataaact
     1261 agacgccttg tcgtcttctt cttcgtattc cttctctttt tcatttttct cctcataaaa
     1321 attaacatag ttattatcgt atccatatat gtatctatcg tatagagtaa attttttgtt
     1381 gtcataaata tatatgtctt ttttaatggg gtgtatagta ccgctgcgca tagtttttct
     1441 gtaatttaca acagtgctat tttctggtag ttcttcggag tgtgttgctt taattattaa
     1501 atttatataa tcaatgaatt tgggatcgtc ggttttgtac aatatgttgc cggcatagta
     1561 cgcagcttct tctagttcaa ttacaccatt ttttagcagc accggattaa cataactttc
     1621 caaaatgttg tacgaaccgt taaacaaaaa cagttcacct cccttttcta tactattgtc
     1681 tgcgagcagt tgtttgttgt taaaaataac agccattgta atgagacgca caaactaata
     1741 tcacaaactg gaaatgtcta tcaatatata gttgctgata tcgggagttc agtcgtcgaa
     1801 tgcaaagcgt aaaaaatatt aataaggtaa aaattacagc tacataaatt acacaattta
     1861 aacggatcca agcttctaga atgcatcacc atcaccatca cgatgacgat gacaagctta
     1921 tggcaggtcc taatcaactc tgcattcgcc gctggactac caagcatgta gctgtgtggc
     1981 tgaaggatga aggctttttt gaatatgtgg acattttatg caataagcac cgacttgatg
     2041 gaatcacatt gctaacattg actgaatatg atctccggtc tcctcctctg gaaatcaaag
     2101 tcttagggga cattaaaagg ttaatgctct cagtccgaaa attgcagaaa atacatattg
     2161 atgttttaga agagatgggc tacaacagtg acagtcccat gggttccatg acccctttca
     2221 tcagtgctct tcagagtaca gactggctct gtaatgggga gctttcccat gactgtgacg
     2281 gacccataac tgacttgaat tctgatcagt accagtacat gaatggtaaa aacaaacatt
     2341 ctgttcgaag attggaccca gaatactgga agactatact gagttgtata tatgttttta
     2401 tagtatttgg atttacatct ttcattatgg ttatagtcca tgagcgagtg cctgacatgc
     2461 agacctatcc accactccca gatatattct tagacagcgt tcctagaatc ccatgggcct
     2521 ttgccatgac ggaagtatgt ggcatgattc tgtgctatat ttggctcctg gttcttcttc
     2581 ttcacaagca caggtcaata cttctgcgaa ggctctgtag tctgatggga actgtattct
     2641 tgcttcgctg ctttaccatg tttgtgacct ccctctccgt gccaggacaa cacctgcagt
     2701 gtactggaaa gatatatggc agtgtatggg agaaattaca tcgagccttt gccatttgga
     2761 gtggctttgg tatgaccctg actggcgttc acacatgtgg agattacatg tttagtggcc
     2821 acacagtcgt cctaactatg ctgaatttct ttgtcaccga atatacacca agaagctgga
     2881 atttcttgca cactttatcc tgggttctca acctctttgg aatcttcttc atcttggctg
     2941 cccatgaaca ttattctatt gatgtgttta ttgcttttta tataacaaca agactctttt
     3001 tgtactacca tactctggcc aataccagag catatcagca gagtaggaga gcaaggattt
     3061 ggtttcccat gttctctttt tttgaatgca atgttaatgg cacagtacct aatgaatatt
     3121 gttggccatt ttcaaaacca gcaataatga aaagactaat tggatgactc gaggtaccgc
     3181 ggccgcagat cttaattaat aaaacacgat acattgttat tagtacattt attaagcgct
     3241 agattctgtg cgttgttgat ttacagacaa ttgttgtacg tattttaata attcattaaa
     3301 tttataatct ttagggtggt atgttagagc gaaaatcaaa tgattttcag cgtctttata
     3361 tctgaattta aatattaaat cctcaataga tttgtaaaat aggtttcgat tagtttcaaa
     3421 caagggttgt ttttccgaac cgatggctgg actatctaat ggattttcgc tcaacgccac
     3481 aaaacttgcc aaatcttgta gcagcaatct agctttgtcg atattcgttt gtgttttgtt
     3541 ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct tttgtatttc tttcatcact
     3601 gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat aacgcttgga catatttaac
     3661 atcgggcgtg ttagctttat taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga
     3721 agttgcttcc gaagacgatt ttgccatagc cacacgacgc ctattaattg tgtcggctaa
     3781 cacgtccgcg atcaaatttg tagttgagct ttttgggcct catgggcctt ccgctcactg
     3841 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca tagctgtttc
     3901 cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3961 gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4441 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4921 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     4981 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5041 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5101 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5161 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5221 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5281 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5341 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5401 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5461 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5521 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5581 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5641 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5761 gcacatttcc ccgaaaagtg ccac
//
