LOCUS       sequence-443738-        6366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses delta 24 alpha synuclein as 
            6XHis-SUMO-eSrtA-LPNTG-Delta-24-ASN fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  AUTHORS   Singh AK, Antonenko A, Kocyla A, Krezel A
  TITLE     An efficient and easily obtainable butelase variant for 
            chemoenzymatic ligation and modification of peptides and proteins.
  JOURNAL   Microb Cell Fact. 2024 Nov 30;23(1):325. doi: 
            10.1186/s12934-024-02598-5.
  PUBMED    39614317
REFERENCE   2  (bases 1 to 6366)
  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: "Microb Cell
            Fact."; date: "2024-11-30"; volume: "23(1):325. doi"; pages: " 
            10.1186/s12934-024-02598-5"
COMMENT     Sequence Label: eSrtA-Delta-24-ASN
FEATURES             Location/Qualifiers
     source          1..6366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..29)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      25..72
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      109..564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(196..215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     406..427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(656..1471)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(656..1471)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1379..1398
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1463..1482
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1593..2181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2082..2101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2335..2352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2367..2506
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2432..2451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2592..2614
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2608..2799)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3608..4690)
                     /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     4652..4671
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4691..4768)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4974..4993
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5077..5096
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5077..5095
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5096..5120
                     /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             5135..5157
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5170..5187
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5218..5238
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             6307..6324
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
       61 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
      121 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      181 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      241 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      301 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
      361 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      421 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
      481 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
      541 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      601 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
      661 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
      721 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
      781 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
      841 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
      901 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
      961 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     1021 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     1081 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     1141 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     1201 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     1261 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     1321 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     1381 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     1441 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     1501 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     1741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2281 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2341 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     2401 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2461 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     2521 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2581 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2641 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     2701 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     2761 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     2821 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     2881 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     2941 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3001 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     3061 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     3121 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     3181 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     3241 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     3301 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     3361 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     3421 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     3481 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     3541 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     3601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     3721 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     3781 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     3841 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     3901 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     3961 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4021 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     4081 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     4141 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     4201 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     4261 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     4321 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     4381 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     4441 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     4501 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     4561 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     4621 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     4681 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     4741 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     4801 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     4861 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     4921 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     4981 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     5041 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat
     5101 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     5161 accatgaaac atcaccatca ccatcacccc atgagcgatt acgacatccc cactactgag
     5221 aatctttatt ttcagggcgc catgggcaac gatcacatta acctgaaagt ggccggtcaa
     5281 gacggtagcg tagtccagtt taaaatcaaa cgccacaccc ctctgtcgaa actgatgaaa
     5341 gcctattgtg aacgccaagg tctgtctatg cgtcagatcc gttttcgctt cgatggacag
     5401 ccgattaacg aaaccgacac tccagcacag ctggaaatgg aagatgagga caccattgac
     5461 gtgttccagc aacagaccgg tggatccatg caagctaaac ctcaaattcc gaaagataaa
     5521 tcaaaagtgg caggctatat tgaaattcca gatgctgata ttaaagaacc agtatatcca
     5581 ggaccagcaa cacgcgaaca attaaataga ggtgtaagct ttgcagaaga aaatgaatca
     5641 ctagatgatc aaaatatttc aattgcagga cacactttca ttgaccgtcc gaactatcaa
     5701 tttacaaatc ttaaagcagc caaaaaaggt agtatggtgt actttaaagt tggtaatgaa
     5761 acacgtaagt ataaaatgac aagtataaga aacgttaagc caacagctgt agaagttcta
     5821 gatgaacaaa aaggtaaaga taaacaatta acattaatta cttgtgatga ttacaatgaa
     5881 gagacaggcg tttgggaaac acgtaaaatc tttgtagcta cagaagtcaa acatatgcta
     5941 ccgaacaccg gtgtggcaga agcagcagga aagacaaaag agggtgttct ctatgtaggc
     6001 tccaaaacca aggagggagt ggtgcatggt gtggcaacag tggctgagaa gaccaaagag
     6061 caagtgacaa atgttggagg agcagtggtg acgggtgtga cagcagtagc ccagaagaca
     6121 gtggagggag cagggagcat tgcagcagcc actggctttg tcaaaaagga ccagttgggc
     6181 aagaatgaag aaggagcccc acaggaagga attctggaag atatgcctgt ggatcctgac
     6241 aatgaggctt atgaaatgcc ttctgaggaa gggtatcaag actacgaacc tgaagcctaa
     6301 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     6361 gagttg
//
