LOCUS       sequence-471035-        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminal His-tagged human SMSr (SAMD8)-sterile alpha motif
            domain deletion mutant protein in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  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 5550)
  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-Delta-SAMD
FEATURES             Location/Qualifiers
     source          1..5550
                     /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           2969..3341
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3693..3710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3803..4391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3883..3902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4562..5353,5354..5422))
                     /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     5185..5204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5423..5527)
                     /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 gacaagcttc
     1921 atattgatgt tttagaagag atgggctaca acagtgacag tcccatgggt tccatgaccc
     1981 ctttcatcag tgctcttcag agtacagact ggctctgtaa tggggagctt tcccatgact
     2041 gtgacggacc cataactgac ttgaattctg atcagtacca gtacatgaat ggtaaaaaca
     2101 aacattctgt tcgaagattg gacccagaat actggaagac tatactgagt tgtatatatg
     2161 tttttatagt atttggattt acatctttca ttatggttat agtccatgag cgagtgcctg
     2221 acatgcagac ctatccacca ctcccagata tattcttaga cagcgttcct agaatcccat
     2281 gggcctttgc catgacggaa gtatgtggca tgattctgtg ctatatttgg ctcctggttc
     2341 ttcttcttca caagcacagg tcaatacttc tgcgaaggct ctgtagtctg atgggaactg
     2401 tattcttgct tcgctgcttt accatgtttg tgacctccct ctccgtgcca ggacaacacc
     2461 tgcagtgtac tggaaagata tatggcagtg tatgggagaa attacatcga gcctttgcca
     2521 tttggagtgg ctttggtatg accctgactg gcgttcacac atgtggagat tacatgttta
     2581 gtggccacac agtcgtccta actatgctga atttctttgt caccgaatat acaccaagaa
     2641 gctggaattt cttgcacact ttatcctggg ttctcaacct ctttggaatc ttcttcatct
     2701 tggctgccca tgaacattat tctattgatg tgtttattgc tttttatata acaacaagac
     2761 tctttttgta ctaccatact ctggccaata ccagagcata tcagcagagt aggagagcaa
     2821 ggatttggtt tcccatgttc tctttttttg aatgcaatgt taatggcaca gtacctaatg
     2881 aatattgttg gccattttca aaaccagcaa taatgaaaag actaattgga tgactcgagg
     2941 taccgcggcc gcagatctta attaataaaa cacgatacat tgttattagt acatttatta
     3001 agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt ttaataattc
     3061 attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc
     3121 tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt ttcgattagt
     3181 ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat tttcgctcaa
     3241 cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat tcgtttgtgt
     3301 tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg tatttctttc
     3361 atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaataacg cttggacata
     3421 tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc
     3481 gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat taattgtgtc
     3541 ggctaacacg tccgcgatca aatttgtagt tgagcttttt gggcctcatg ggccttccgc
     3601 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc
     3661 tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc
     3721 gttcgggtaa agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta
     3781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4681 ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa
     4741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5401 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5521 ttccgcgcac atttccccga aaagtgccac
//
