LOCUS       sequence-353207-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in bacterial cells. PDZ1 and PDZ2 domains 
            (truncated at C-terminus), G111-F275..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5853)
  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            "unpublished"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-c021
FEATURES             Location/Qualifiers
     source          1..5853
                     /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             671..688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(741..759)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      755..802
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      839..1294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(926..945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1136..1157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1386..2201)
                     /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(1386..2201)
                     /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     2109..2128
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2193..2212
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2323..2911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2812..2831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3065..3082
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3097..3236
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3162..3181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3322..3344
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3338..3529)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4338..5420)
                     /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     5382..5401
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5421..5498)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5704..5723
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5807..5826
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5807..5825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5826..5850
                     /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 tcttttgaca gtaaaggtgg atacggatcc gaattcgagc tccgtcgaca agcttgcggc
      661 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
      721 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      781 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
      841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1201 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1261 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1321 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     1381 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1441 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1501 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1561 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1621 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1681 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1741 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1801 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1861 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1921 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1981 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2041 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2101 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2161 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     2221 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     2281 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2341 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2401 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2461 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2521 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2581 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2641 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2701 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2761 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2821 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2881 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2941 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3001 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3061 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3121 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     3181 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     3241 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3301 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3361 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3421 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3481 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3541 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     3601 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     3661 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     3721 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     3781 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     3841 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3901 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3961 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     4021 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     4081 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     4141 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     4201 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     4261 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     4321 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4381 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     4441 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     4501 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4561 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     4621 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4681 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4741 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4801 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4861 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4921 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4981 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     5041 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     5101 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     5161 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     5221 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     5281 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     5341 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     5401 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     5461 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     5521 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     5581 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     5641 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     5701 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     5761 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     5821 ataggggaat tgtgagcgga taacaattcc cct
//
