LOCUS       sequence-482093-        5748 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Expresses alpha synuclein (missing exon 3, residues 41-54) in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5748)
  AUTHORS   SanGiovanni DQ, McGlinchey RP, Lee JC
  TITLE     Amyloid formation of alternatively spliced variants of 
            alpha-synuclein.
  JOURNAL   Protein Sci. 2025 Jul;34(7):e70195. doi: 10.1002/pro.70195.
  PUBMED    40522183
REFERENCE   2  (bases 1 to 5748)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2025-07"; volume: "34(7):e70195. doi"; pages: " 
            10.1002/pro.70195"
COMMENT     Sequence Label: pET21a asyn(Delta-41-54)
FEATURES             Location/Qualifiers
     source          1..5748
                     /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             476..493
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(546..564)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      560..607
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      644..1099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(731..750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     941..962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1126..1230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1231..1299,1300..2091)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1449..1468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2262..2850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2751..2770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3004..3021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3036..3178
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3104..3123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3264..3286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3280..3471)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4280..5362)
                     /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     5324..5343
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5363..5440)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5646..5665
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggatgtattc atgaaaggac tttcaaaggc
      121 caaggaggga gttgtggctg ctgctgagaa aaccaaacag ggtgtggcag aagcagcagg
      181 aaagacaaaa gagggtgttc tctatgtagt ggctgagaag accaaagagc aagtgacaaa
      241 tgttggagga gcagtggtga cgggtgtgac agcagtagcc cagaagacag tggagggagc
      301 agggagcatt gcagcagcca ctggctttgt caaaaaggac cagttgggca agaatgaaga
      361 aggagcccca caggaaggaa ttctggaaga tatgcctgtg gatcctgaca atgaggctta
      421 tgaaatgcct tctgaggaag ggtatcaaga ctatgaacct gaagcctaac tcgagcacca
      481 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      541 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
      601 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
      661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      841 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      901 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      961 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1021 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1081 atattaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1141 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1201 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1261 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1321 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1381 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1441 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1501 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1561 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1621 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1681 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1741 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     1801 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1861 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1921 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1981 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2041 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2101 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2401 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3001 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     3121 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3181 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3241 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3301 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3361 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3421 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     3481 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     3541 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     3601 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     3661 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     3721 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     3781 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3841 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3901 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     3961 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4021 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4081 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4141 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4201 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4261 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4381 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4441 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4501 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4561 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4621 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4681 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     4741 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     4801 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     4861 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     4921 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     4981 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5041 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5101 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     5161 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     5221 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5281 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     5341 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     5401 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5461 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5521 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     5581 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     5641 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     5701 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
