LOCUS       sequence-384746-        8095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Purification of SARM1 from insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8095)
  TITLE     Wu Lab 2022
REFERENCE   2  (bases 1 to 8095)
  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"
COMMENT     Sequence Label: pFastBac HTB MBP-SARM1
FEATURES             Location/Qualifiers
     source          1..8095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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     complement(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(3915..4024)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4048..4139
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4064..4081
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4193..4210
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4214..5311
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      5168..5177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5285..5308
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    5330..5354
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5354..5374
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(7416..7440)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7585..7719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7622..7641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7676..7695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7748..7913
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg cccttgcttc aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta ctcgagatcc cgggtgatca
     3901 agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat
     3961 acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa
     4021 ggtccgtata ctccggaata ttaatagatc atggagataa ttaaaatgat aaccatctcg
     4081 caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata
     4141 ttccggatta ttcataccgt cccaccatcg ggcgcggatc accatgggat cccatcacca
     4201 tcaccatcac ggcaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg
     4261 ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac
     4321 cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg
     4381 ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt
     4441 ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc
     4501 cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat
     4561 ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga
     4621 taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt
     4681 cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta
     4741 cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt
     4801 tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc
     4861 ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga
     4921 caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa
     4981 accgttcgtt ggcgtgctga gcgcaggtat taacgctgcc agtccgaaca aagagctggc
     5041 aaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga
     5101 caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg
     5161 tattgccgcc accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat
     5221 gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac
     5281 tgtcgatgaa gccctgaaag acgcgcagac taattcggga tcctcgatca caagtttgta
     5341 caaaaaagtt ggcgaaaacc tgtacttcca aggcccacgg ccgggcgccg agcggctggc
     5401 ggtgcctggg ccagatgggg gcggtggcac gggcccatgg tgggctgcgg gtggccgcgg
     5461 gccccgcgaa gtgtcgccgg gggcaggcac cgaggtgcag gacgccctgg agcgcgcgct
     5521 gccggagctg cagcaggcct tgtccgcgct gaagcaggcg ggcggcgcgc gggccgtggg
     5581 cgccggcctg gccgaggtct tccaactggt ggaggaggcc tggctgctgc cggccgtggg
     5641 ccgcgaggta gcccagggtc tgtgcgacgc catccgcctc gatggcggcc tcgacctgct
     5701 gttgcggctg ctgcaggcgc cggagttgga gacgcgtgtg caggccgcgc gcctgctgga
     5761 gcagatcctg gtggctgaga accgagaccg cgtggcgcgc attgggctgg gcgtgatcct
     5821 gaacctggcg aaggaacgcg aacccgtaga gctggcgcgg agcgtggcag gcatcttgga
     5881 gcacatgttc aagcattcgg aggagacatg ccagaggctg gtggcggccg gcggcctgga
     5941 cgcggtgctg tattggtgcc gccgcacgga ccccgcgctg ctgcgccact gcgcgctggc
     6001 gctgggcaac tgcgcgctgc acgggggcca ggcggtgcag cgacgcatgg tagagaagcg
     6061 cgcagccgag tggctcttcc cgctcgcctt ctccaaggag gacgagctgc ttcggctgca
     6121 cgcctgcctc gcagtagcgg tgttggcgac taacaaggag gtggagcgcg aggtggagcg
     6181 ctcgggcacg ctggcgctcg tggagccgct tgtggcctcg ctggaccctg gccgcttcgc
     6241 ccgctgtctg gtggacgcca gcgacacaag ccagggccgc gggcccgacg acctgcagcg
     6301 cctcgtgccg ttgctcgact ctaaccgctt ggaggcgcag tgcatcgggg ctttctacct
     6361 ctgcgccgag gctgccatca agagcctgca aggcaagacc aaggtgttca gcgacatcgg
     6421 cgccatccag agcctgaaac gcctggtttc ctactctacc aatggcacta agtcggcgct
     6481 ggccaagcgc gcgctgcgcc tgctgggcga ggaggtgcca cggcccatcc tgccctccgt
     6541 gcccagctgg aaggaggccg aggttcagac gtggctgcag cagatcggtt tctccaagta
     6601 ctgcgagagc ttccgggagc agcaggtgga tggcgacctg cttctgcggc tcacggagga
     6661 ggaactccag accgacctgg gcatgaaatc gggcatcacc cgcaagaggt tctttaggga
     6721 gctcacggag ctcaagacct tcgccaacta ttctacgtgc gaccgcagca acctggcgga
     6781 ctggctgggc agcctggacc cgcgcttccg ccagtacacc tacggcctgg tcagctgcgg
     6841 cctggaccgc tccctgctgc accgcgtgtc tgagcagcag ctgctggaag actgcggcat
     6901 ccacctgggc gtgcaccgcg cccgcatcct cacggcggcc agagaaatgc tacactcccc
     6961 gctgccctgt actggtggca aacccagtgg ggacactcca gatgtcttca tcagctaccg
     7021 ccggaactca ggttcccagc tggccagtct cctgaaggtg cacctgcagc tgcatggctt
     7081 cagtgtcttc attgatgtgg agaagctgga agcaggcaag ttcgaggaca aactcatcca
     7141 gagtgtcatg ggtgcccgca actttgtgtt ggtgctatca cctggagcac tggacaagtg
     7201 catgcaagac catgactgca aggattgggt gcataaggag attgtgactg ctttaagctg
     7261 cggcaagaac attgtgccca tcattgatgg cttcgagtgg cctgagcccc aggtcctgcc
     7321 tgaggacatg caggctgtgc ttactttcaa cggtatcaag tggtcccacg aataccagga
     7381 ggccaccatt gagaagatca tccgcttcct gcagtaatca actttcttgt acaaagtggt
     7441 gatcgagcta gctagaagct tgtcgagaag tactagagga tcataatcag ccataccaca
     7501 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     7561 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     7621 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7681 ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct tgagcctagg
     7741 agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     7801 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     7861 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     7921 atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg acaaaccgtc
     7981 atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct cttcgttatt
     8041 aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg aatgg
//
