LOCUS       sequence-482094-        5706 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Expresses alpha synuclein (missing exon 5, residues 103-130) in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5706)
  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 5706)
  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-103-130)
FEATURES             Location/Qualifiers
     source          1..5706
                     /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             434..451
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(504..522)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      518..565
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      602..1057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(689..708)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     899..920
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1084..1188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1189..1257,1258..2049)
                     /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(1407..1426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2220..2808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2709..2728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2962..2979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2994..3136
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3062..3081)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3222..3244
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3238..3429)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4238..5320)
                     /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     5282..5301
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5321..5398)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5604..5623
                     /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 tctatgtagg ctccaaaacc aaggagggag tggtgcatgg
      241 tgtggcaaca gtggctgaga agaccaaaga gcaagtgaca aatgttggag gagcagtggt
      301 gacgggtgtg acagcagtag cccagaagac agtggaggga gcagggagca ttgcagcagc
      361 cactggcttt gtcaaaaagg accagttggg caaggaaggg tatcaagact atgaacctga
      421 agcctaactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
      481 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      541 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
      601 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      661 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      721 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      781 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      841 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      901 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      961 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1021 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
     1081 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1141 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1201 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1261 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1321 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1381 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1441 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1501 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1561 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1621 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1681 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     1741 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1801 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1861 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1921 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1981 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2041 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2341 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca
     3061 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3121 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3181 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3241 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3301 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3361 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3421 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3481 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3541 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3601 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     3661 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     3721 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     3781 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     3841 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     3901 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     3961 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4021 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4081 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     4141 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4201 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4261 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4321 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4381 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4441 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4501 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4561 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4621 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4681 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4741 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     4801 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     4861 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     4921 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     4981 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5041 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5101 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5161 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5221 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5281 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5341 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5401 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5461 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     5521 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     5581 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     5641 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     5701 cgaaat
//
