LOCUS       sequence-460633-        5725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of 21 kDa mouse proSAAS chaperone protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5725)
  AUTHORS   Hoshino A, Helwig M, Rezaei S, Berridge C, Eriksen JL, Lindberg I
  TITLE     A novel function for proSAAS as an amyloid anti-aggregant in 
            Alzheimer's disease.
  JOURNAL   J Neurochem. 2014 Feb;128(3):419-30. doi: 10.1111/jnc.12454. Epub 
            2013 Oct 24.
  PUBMED    24102330
REFERENCE   2  (bases 1 to 5725)
  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: "10.1111/jnc.12454";
            journalName: "J Neurochem"; date: "2014-02"; volume: "128"; issue: 
            "3"; pages: "419-30"
COMMENT     Sequence Label: pET45 His-proSAAS (21 kDa)
FEATURES             Location/Qualifiers
     source          1..5725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     477..496
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(516..593)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     799..818
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     906..925
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        906..924
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    925..949
                     /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             964..986
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1002..1019
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1596..1640
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1678..1696)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1692..1739
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1776..2231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1863..1882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2073..2094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2346..3137,3138..3206))
                     /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     2969..2988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3207..3299)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3380..3968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3869..3888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4122..4139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4154..4293
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4219..4238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4379..4401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4395..4586)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5158..5725,1..515))
                     /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"
ORIGIN
        1 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
       61 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
      121 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
      181 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
      241 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
      301 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
      361 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
      421 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
      481 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
      541 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
      601 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
      661 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
      721 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
      781 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
      841 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc
      901 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
      961 aattttgttt aactttaaga aggagatata ccatggggat gcaccaccac caccaccacg
     1021 gcagcgcgag gcccgtgaag gagccccgca gtctgagcgc agcatccgcg cccttggttg
     1081 agacgagcac tcccctccgc ttgcgtcggg ccgtgccccg aggagaggcg gcgggtgcgg
     1141 tgcaggagct ggcgcgggcg ctggcgcacc tgctggaggc cgagagacag gaacgcgcgc
     1201 gtgctgaggc gcaggaggct gaggatcagc aggcgcgtgt cctggcgcag ctgctgcgcg
     1261 cctggggctc tccgcgtgcc tcggacccgc ccttggcccc cgacgatgac ccggacgctc
     1321 cagctgcaca gctcgcccgt gctctgctcc gagctcgcct agacccggcc gccctggcag
     1381 cccaacttgt ccccgcccct gccgctgcgc cgcgaccccg gcccccagtg tatgatgatg
     1441 gccccactgg cccagacgtc gaggatgccg gcgacgagac tcctgacgtg gaccctgagc
     1501 tgctgaggta cttgctaggg cggatcctca ccggaagttc ggagccagag gctgctcctg
     1561 ccccgtaaaa gcttgcggcc gcactcgagt ctggtaaaga aaccgctgct gcgaaatttg
     1621 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
     1681 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1741 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     1801 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1861 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1921 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1981 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2041 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2101 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2161 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2221 gtttacaatt tctggcggca cgatggcatg agattatcaa aaaggatctt cacctagatc
     2281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatcatgatt gaagcattta
     3241 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3301 aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3361 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3421 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3481 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3541 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3601 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3661 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3721 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3781 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3841 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3901 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3961 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4021 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4081 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4141 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4201 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     4261 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4321 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4381 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     4441 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     4501 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     4561 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     4621 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     4681 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     4741 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     4801 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     4861 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4921 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4981 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     5041 atgctagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     5101 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     5161 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5221 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     5281 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     5341 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     5401 tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc
     5461 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     5521 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     5581 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     5641 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     5701 ctggtgaccc aatgcgacca gatgc
//
