LOCUS       sequence-174096-        4433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of rat Alpha-synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4433)
  TITLE     MJFF plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4433)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pD454-SR rat Alpha-synuclein
FEATURES             Location/Qualifiers
     source          1..4433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      444..491
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(714..733)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(739..758)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        800..891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(892..960,961..1752)
                     /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     complement(1110..1129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1835..2917
                     /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     complement(1854..1873)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3003..3591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3492..3511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3745..3762
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3860..3877
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3861..3877
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4340..4359
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4340..4358
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4359..4383
                     /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             4420..4428
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 atggacgtgt ttatgaaagg tctgagcaag gcaaaggaag gtgtagtggc agcggcggag
       61 aaaacgaaac agggcgtggc ggaagcggcg ggtaagacga aagagggcgt cctgtatgtt
      121 ggtagcaaga ccaaggaagg cgtggtccac ggtgttacca ccgttgctga gaaaactaaa
      181 gaacaagtta ccaacgtcgg tggcgccgtg gttacgggcg tgaccgcagt tgcgcaaaag
      241 accgtcgagg gtgcgggtaa tatcgccgct gcgaccggtt tcgtcaaaaa agaccagatg
      301 ggtaagggtg aggaaggcta cccgcaagag ggcattctgg aagatatgcc ggtggacccg
      361 agctccgagg catacgaaat gccgagcgaa gaaggctatc aggattacga gccagaggcc
      421 taatgaggtt gaggtctcac cccctagcat aaccccttgg ggcctctaaa cgggtcttga
      481 ggggtttttt gcccctgaga cgcgtcaatc gagttcgtac ctaagggcga caccccataa
      541 ttagcccggg cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc
      601 agttccctac tctcgcatgg ggagtcccca cactaccatc ggcgctacgg cgtttcactt
      661 ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc aaacaagggg
      721 tgttatgagc catattcagg tataaatggg ctcgcgataa tgttcagaat tggttaattg
      781 gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa tatgagtatt
      901 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      961 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1021 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1081 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1141 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1201 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1261 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1321 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1381 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagcg
     1441 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1501 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1561 ccggctggct ggtttattgc tgataaatcc ggagccggtg agcgtggttc tcgcggtatc
     1621 atcgcagcgc tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1681 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1741 aagcattggt aagcggcgcg ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     1801 gcgcccggaa gagagtcaat tcagggtggt gaatatgaaa ccagtaacgt tatacgatgt
     1861 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     1921 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     1981 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     2041 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     2101 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     2161 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     2221 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     2281 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaggacg gtacgcgact
     2341 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     2401 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     2461 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     2521 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     2581 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     2641 ggtagtggga tacgacgata ccgaagatag ctcatgttat atcccgccgt taaccaccat
     2701 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     2761 gggccaggcg gtgaagggca atcagctgtt gccagtctca ctggtgaaaa gaaaaaccac
     2821 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2881 ggcacgacag gtttcccgac tggaaagcgg gcagtgactc atgaccaaaa tcccttaacg
     2941 tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt agcccaccac
     3181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt
     3781 agggaactgc caggcatcaa actaagcaga aggcccctga cggatggcct ttttgcgttt
     3841 ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc cccctgggcg
     3901 gttctgataa cgagtaatcg ttaatccgca aataacgtaa aaacccgctt cggcgggttt
     3961 ttttatgggg ggagtttagg gaaagagcat ttgtcagaat atttaagggc gcctgtcact
     4021 ttgcttgata tatgagaatt atttaacctt ataaatgaga aaaaagcaac gcactttaaa
     4081 taagatacgt tgctttttcg attgatgaac acctataatt aaactattca tctattattt
     4141 atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa aataaatctc
     4201 gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca acgataatac
     4261 aaaatataat acaaactata agatgttatc agtatttatt atcatttaga ataaattttg
     4321 tgtcgccctt ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
     4381 tcccctctag aaataatttt gtttaacttt ttgagacctt aaggaggtaa aaa
//
