LOCUS       sequence-353208-        5922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in bacterial cells. PDZ1 and PDZ2 domains, 
            G111-V288..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5922)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5922)
  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            "unpublished"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-c020
FEATURES             Location/Qualifiers
     source          1..5922
                     /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             740..757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(810..828)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      824..871
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      908..1363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(995..1014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1205..1226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1455..2270)
                     /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(1455..2270)
                     /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     2178..2197
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2262..2281
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2392..2980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2881..2900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3134..3151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3166..3305
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3231..3250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3391..3413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3407..3598)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4407..5489)
                     /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     5451..5470
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5490..5567)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5773..5792
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5876..5895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5876..5894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5895..5919
                     /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 gggattcgtg
      121 aagtcatttt gtgtaaggat caagatggaa aaattggact caggcttaaa tcaatagata
      181 atggtatatt tgttcagcta gtccaggcta attctccagc ctcattggtt ggtctgagat
      241 ttggggacca agtacttcag atcaatggtg aaaactgtgc aggatggagc tctgataaag
      301 cgcacaaggt gctcaaacag gcttttggag agaagattac catgaccatt cgtgacaggc
      361 cctttgaacg gacgattacc atgcataagg atagcactgg acatgttggt tttatcttta
      421 aaaatggaaa aataacatcc atagtgaaag atagctctgc agccagaaat ggtcttctca
      481 cggaacataa catctgtgaa atcaatggac agaatgtcat tggattgaag gactctcaaa
      541 ttgcagacat actgtcaaca tctgggactg tagttactat tacaatcatg cctgctttta
      601 tctttgaaca tattattaag cggatggcac caagcattat gaaaagccta atggaccaca
      661 ccattcctga ggtttgacag taaaggtgga tacggatccg aattcgagct ccgtcgacaa
      721 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
      781 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      841 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
      901 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      961 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1021 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1081 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1141 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1201 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1261 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1321 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     1381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1441 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     1501 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     1561 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     1621 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     1681 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     1741 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     1801 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     1861 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     1921 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     1981 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     2041 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     2101 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     2161 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     2221 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     2281 gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga
     2341 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2521 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3061 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3121 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3181 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3241 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3301 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3361 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3421 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3481 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3541 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3601 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3661 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3721 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     3781 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     3841 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     3901 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     3961 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4021 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     4081 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     4141 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     4201 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     4261 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     4321 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     4381 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4441 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     4501 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     4561 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     4621 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     4681 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     4741 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     4801 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     4861 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     4921 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     4981 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     5041 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     5101 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     5161 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     5221 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     5281 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     5341 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     5401 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     5461 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     5521 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     5581 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     5641 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     5701 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     5761 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     5821 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
     5881 cgactcacta taggggaatt gtgagcggat aacaattccc ct
//
