LOCUS       sequence-336172-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Saposin B (Homo Sapien) containing a single mutation, arginine 38 to
            tryptophan.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  AUTHORS   Kurgan KW, Chen B, Brown KA, Falco Cobra P, Ye X, Ge Y, Gellman SH
  TITLE     Stable Picodisc Assemblies from Saposin Proteins and Branched 
            Detergents.
  JOURNAL   Biochemistry. 2021 Apr 13;60(14):1108-1119. doi: 
            10.1021/acs.biochem.0c00924. Epub 2021 Mar 23.
  PUBMED    33755420
REFERENCE   2  (bases 1 to 5942)
  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: 
            "10.1021/acs.biochem.0c00924"; journalName: "Biochemistry"; date: 
            "2021-04-13- 13"; volume: "60"; issue: "14"; pages: "1108-1119"
COMMENT     Sequence Label: saposin B^W
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(443..461)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      457..504
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(679..684,685..701,702..707))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(734..752)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        820..924
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(925..993,994..1785)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1143..1162)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1956..2544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2445..2464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2698..2715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2730..2869
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2795..2814)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2955..2977
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2971..3162)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4474..5556)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5518..5537
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5557..5634)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5840..5859
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg ggtgacgttt gccaagattg tattcaaatg
      181 gttaccgata ttcagaccgc cgttcgtacc aattctacct ttgttcaagc cttggttgaa
      241 catgttaaag aagaatgtga ttggttgggt cctggtatgg ccgatatttg caaaaattat
      301 attagccagt atagcgaaat cgctattcag atgatgatgc atatgcaacc gaaagaaatt
      361 tgtgctttgg ttggtttttg tgattaactc gaggatccgg ctgctaacaa agcccgaaag
      421 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
      481 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga
      541 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga
      601 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta
      661 actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct
      721 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      781 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      841 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      901 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      961 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1021 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1081 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1141 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     1201 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1261 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1321 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1381 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1441 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     1501 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1561 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1621 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1681 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1741 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1801 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1861 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1921 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1981 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2041 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2101 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2161 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2221 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2281 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2341 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2401 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2461 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2521 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2581 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2641 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2701 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     2761 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2821 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     2881 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2941 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3001 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     3061 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     3121 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     3181 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     3241 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     3301 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     3361 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     3421 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     3481 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     3541 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     3601 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg
     3661 cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc
     3721 attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc
     3781 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
     3841 gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg
     3901 ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg
     3961 gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac
     4021 agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg
     4081 aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc
     4141 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     4201 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     4261 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     4321 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     4381 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4441 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4501 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4561 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4621 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4681 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4741 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4801 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4861 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4921 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4981 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5041 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     5101 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     5161 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     5221 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     5281 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5341 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5401 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5461 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5521 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5581 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5641 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5701 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5761 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5821 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5881 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     5941 at
//
