LOCUS       sequence-482095-        5664 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Expresses alpha synuclein (missing exon 3 and 5, residues 41-54 and 
            103-130) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  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 5664)
  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/Delta-103-130)
FEATURES             Location/Qualifiers
     source          1..5664
                     /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             392..409
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(462..480)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      476..523
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      560..1015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(647..666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     857..878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1042..1146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1147..1215,1216..2007)
                     /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(1365..1384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2178..2766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2667..2686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2920..2937
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2952..3094
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3020..3039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3180..3202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3196..3387)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4196..5278)
                     /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     5240..5259
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5279..5356)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5562..5581
                     /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 aggaagggta
      361 tcaagactat gaacctgaag cctaactcga gcaccaccac caccaccact gagatccggc
      421 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      481 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      541 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      601 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      661 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      721 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      781 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      841 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      901 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      961 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcaggt
     1021 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1081 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1141 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1201 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1261 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1321 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1381 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1441 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1501 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1561 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1621 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1681 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1741 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1801 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1861 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1921 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1981 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2041 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2101 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2221 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2341 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2821 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2881 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2941 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3001 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3061 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3121 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3181 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3241 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3301 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3361 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3421 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3481 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3541 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     3601 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     3661 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     3721 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     3781 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     3841 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     3901 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     3961 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     4021 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     4081 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     4141 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4261 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4321 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4381 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     4441 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     4501 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     4561 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     4621 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     4681 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     4741 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     4801 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     4861 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     4921 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     4981 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     5041 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     5101 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     5161 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     5221 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     5281 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     5341 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     5401 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     5461 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     5521 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     5581 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     5641 atcgagatct cgatcccgcg aaat
//
