LOCUS       sequence-353180-        6038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in bacterial cells. Entire CDS, M1-K199. Can be 
            biotinylated..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6038)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6038)
  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            "unpublished"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: PRDX1A-c001
FEATURES             Location/Qualifiers
     source          1..6038
                     /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             759..803
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             856..873
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(926..944)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      940..987
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1024..1479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1111..1130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1321..1342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1571..2386)
                     /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(1571..2386)
                     /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     2294..2313
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2378..2397
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2508..3096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2997..3016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3250..3267
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3282..3421
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3347..3366)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3507..3529
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3523..3714)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4523..5605)
                     /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     5567..5586
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5606..5683)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5889..5908
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5992..6011
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5992..6010
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6011..6035
                     /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 tcttcaggaa
      121 atgctaaaat tgggcaccct gcccccaact tcaaagccac agctgttatg ccagatggtc
      181 agtttaaaga tatcagcctg tctgactaca aaggaaaata tgttgtgttc ttcttttacc
      241 ctcttgactt cacctttgtg tgccccacgg agatcattgc tttcagtgat agggcagaag
      301 aatttaagaa actcaactgc caagtgattg gtgcttctgt ggattctcac ttctgtcatc
      361 tagcatgggt caatacacct aagaaacaag gaggactggg acccatgaac attcctttgg
      421 tatcagaccc gaagcgcacc attgctcagg attatggggt cttaaaggct gatgaaggca
      481 tctcgttcag gggccttttt atcattgatg ataagggtat tcttcggcag atcactgtaa
      541 atgacctccc tgttggccgc tctgtggatg agactttgag actagttcag gccttccagt
      601 tcactgacaa acatggggaa gtgtgcccag ctggctggaa acctggcagt gataccatca
      661 agcctgatgt ccaaaagagc aaagaatatt tctccaagca gaagagcagt aaaggtggat
      721 acggctctgg aggcggtgga agtggtggcg gaggtagcgg tctgaatgat atctttgaag
      781 cgcagaagat tgaatggcat gaatgataag gatccgaatt cgagctccgt cgacaagctt
      841 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
      901 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      961 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     1021 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1081 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1141 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1201 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1261 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1321 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1381 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1441 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     1501 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1561 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1621 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1681 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1741 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1801 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     1861 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1921 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1981 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     2041 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     2101 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     2161 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     2221 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     2281 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     2341 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     2401 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
     2461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2641 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2701 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2761 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2821 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2881 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2941 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3001 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3061 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3121 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3181 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3241 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3301 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     3361 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     3421 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3481 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3541 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3601 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3661 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     3721 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     3781 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3841 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     3901 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     3961 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     4021 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     4081 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     4141 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat
     4201 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     4261 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     4321 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     4381 aaagaagaca gtcataagtg cggcgacgat agtcatgtcc cgcgcccacc ggaaggagct
     4441 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     4501 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     4621 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     4681 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     4741 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     4801 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     4861 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4921 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4981 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     5041 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     5101 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     5161 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     5221 ccagctgata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     5281 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     5341 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     5401 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     5461 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     5521 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     5581 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     5641 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     5701 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     5761 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     5821 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     5881 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     5941 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
     6001 tcactatagg ggaattgtga gcggataaca attcccct
//
