LOCUS       sequence-353168-        5937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in bacterial cells. PDZ1 and PDZ2 domains, 
            A106-V288. Can be used for crystallography..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5937)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5937)
  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            "unpublished"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: SDCBPA-c010
FEATURES             Location/Qualifiers
     source          1..5937
                     /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             755..772
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(825..843)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      839..886
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      923..1378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1010..1029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1220..1241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1470..2285)
                     /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(1470..2285)
                     /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     2193..2212
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2277..2296
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2407..2995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2896..2915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3149..3166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3181..3320
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3246..3265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3406..3428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3422..3613)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4422..5504)
                     /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     5466..5485
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5505..5582)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5788..5807
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5891..5910
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5891..5909
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5910..5934
                     /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 gcagaaatta
      121 agcaagggat tcgtgaagtc attttgtgta aggatcaaga tggaaaaatt ggactcaggc
      181 ttaaatcaat agataatggt atatttgttc agctagtcca ggctaattct ccagcctcat
      241 tggttggtct gagatttggg gaccaagtac ttcagatcaa tggtgaaaac tgtgcaggat
      301 ggagctctga taaagcgcac aaggtgctca aacaggcttt tggagagaag attaccatga
      361 ccattcgtga caggcccttt gaacggacga ttaccatgca taaggatagc actggacatg
      421 ttggttttat ctttaaaaat ggaaaaataa catccatagt gaaagatagc tctgcagcca
      481 gaaatggtct tctcacggaa cataacatct gtgaaatcaa tggacagaat gtcattggat
      541 tgaaggactc tcaaattgca gacatactgt caacatctgg gactgtagtt actattacaa
      601 tcatgcctgc ttttatcttt gaacatatta ttaagcggat ggcaccaagc attatgaaaa
      661 gcctaatgga ccacaccatt cctgaggttt gacagtaaag gtggatacgg atccgaattc
      721 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
      781 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      841 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      901 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      961 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1021 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1081 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1141 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1201 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1261 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1321 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1381 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1441 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     1501 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1561 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1621 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1681 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     1741 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1801 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1861 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1921 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1981 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     2041 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     2101 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     2161 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2221 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     2281 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     2341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2521 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3181 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3241 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3301 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3361 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3421 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3481 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3541 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3601 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3661 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3721 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3781 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     3841 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     3901 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     3961 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4021 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4081 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4141 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4201 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4261 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4321 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4381 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4441 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4501 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4561 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4621 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     4681 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     4741 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     4801 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     4861 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     4921 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     4981 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5041 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5101 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5161 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5221 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5281 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5341 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5401 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5461 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5521 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     5581 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     5641 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     5701 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     5761 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     5821 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     5881 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccct
//
