LOCUS       sequence-353175-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in bacterial cells. Entire CDS, M1-P224. Can be 
            biotinylated..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6113)
  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            "unpublished"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: PRDX6A-c001
FEATURES             Location/Qualifiers
     source          1..6113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             12..34
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             45..62
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             87..107
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             834..878
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             931..948
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1001..1019)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1015..1062
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1099..1554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1186..1205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1396..1417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1646..2461)
                     /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(1646..2461)
                     /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     2369..2388
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2453..2472
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2583..3171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3072..3091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3325..3342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3357..3496
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3422..3441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3582..3604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3598..3789)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4598..5680)
                     /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
                     EACKAAVHNLLAQRVSGLIINYQLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5642..5661
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5681..5758)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5964..5983
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6067..6086
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6067..6085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6086..6110
                     /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)."
ORIGIN
        1 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccat catcatcatc
       61 attcttctgg tgtagatctg ggtaccgaga acctgtactt ccaatccatg cccggaggtc
      121 tgcttctcgg ggacgtggct cccaactttg aggccaatac caccgtcggc cgcatccgtt
      181 tccacgactt tctgggagac tcatggggca ttctcttctc ccaccctcgg gactttaccc
      241 cagtgtgcac cacagagctt ggcagagctg caaagctggc accagaattt gccaagagga
      301 atgttaagtt gattgccctt tcaatagaca gtgttgagga ccatcttgcc tggagcaagg
      361 atatcaatgc ttacaattgt gaagagccca cagaaaagtt accttttccc atcatcgatg
      421 ataggaatcg ggagcttacc atcctgttgg gcatgctgga tccagcagag aaggatgaaa
      481 agggcatgcc tgtgacagct cgtgtggtgt ttgtttttgg tcctgataag aagctgaagc
      541 tgtctatcct ctacccagct accactggca ggaactttga tgagattctc agggtagtca
      601 tctctctcca gctgacagca gaaaaaaggg ttgccacccc agttgattgg aaggatgggg
      661 atagtgtgat ggtccttcca accatccctg aagaagaagc caaaaaactt ttcccgaaag
      721 gagtcttcac caaagagctc ccatctggca agaaatacct ccgctacaca ccccagccta
      781 gcagtaaagg tggatacggc tctggaggcg gtggaagtgg tggcggaggt agcggtctga
      841 atgatatctt tgaagcgcag aagattgaat ggcatgaatg ataaggatcc gaattcgagc
      901 tccgtcgaca agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct
      961 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     1021 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     1081 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1141 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1441 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     1561 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1621 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     1681 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1741 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1801 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1861 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     1921 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1981 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     2041 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     2101 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     2161 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     2221 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2281 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     2341 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2401 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     2461 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc
     2521 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2581 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2641 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2701 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2761 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2821 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2881 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2941 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3001 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3061 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3241 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3301 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3361 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     3421 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3481 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3541 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3601 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3661 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3721 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3781 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3841 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3901 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3961 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     4021 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     4081 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     4141 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4201 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     4261 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4321 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4381 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4441 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgtcccgcgc
     4501 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4561 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4621 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4681 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4741 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4801 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4861 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4921 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4981 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     5041 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     5101 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     5161 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     5221 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5281 tggcatcctg gtcatccagc tgatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5341 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5401 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5461 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5521 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5581 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5641 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5701 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5761 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5821 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     5881 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     5941 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6001 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     6061 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cct
//
