LOCUS       sequence-353150-        5827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in bacterial cells (as inclusion bodies). CD44 
            Link module, M63-N178. Used for crystallography..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5827)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5827)
  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            "unpublished"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: CD44I-c011
FEATURES             Location/Qualifiers
     source          1..5827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             580..600
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             601..618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             628..642
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             692..709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(762..780)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      776..823
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      860..1315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(947..966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1157..1178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1407..2222)
                     /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(1407..2222)
                     /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     2130..2149
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2214..2233
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2344..2932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2833..2852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3086..3103
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3118..3257
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3183..3202)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3343..3365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3359..3550)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4359..5441)
                     /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     5403..5422
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5442..5519)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5725..5744
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaacct taagaaggag atatactatg gctatggcgc agatcgattt gaatataacc
      121 tgccgctttg caggtgtatt ccacgtggag aaaaatggtc gctacagcat ctctcggacg
      181 gaggccgctg acctctgcaa ggctttcaat agcaccttgc ccacaatggc ccagatggag
      241 aaagctctga gcatcggatt tgagacctgc aggtatgggt tcatagaagg gcatgtggtg
      301 attccccgga tccaccccaa ctccatctgt gcagcaaaca acacaggggt gtacatcctc
      361 acatccaaca cctcccagta tgacacatat tgcttcaatg cttcagctcc acctgaagaa
      421 gattgtacat cagtcacaga cctgcccaat gcctttgatg gaccaattac cataactatt
      481 gttaaccgtg atggcacccg ctatgtccag aaaggagaat acagaacgaa tcctgaagac
      541 atctacccca gcaaccctac tgatgatgac gtgtgagcag agaacctcta cttccaatcg
      601 caccatcatc accaccattg atacaaggat gacgacgata agtgaggatc cgaattcgag
      661 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
      721 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      781 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      841 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      901 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      961 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1021 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1081 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1141 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1201 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1261 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     1321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1381 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     1441 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     1501 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     1561 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     1621 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     1681 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     1741 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     1801 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     1861 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     1921 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     1981 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     2041 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     2101 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2161 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     2221 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
     2281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3001 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3061 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3121 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     3181 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3241 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3301 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3361 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3421 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3481 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3541 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     3601 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     3661 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     3721 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     3781 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     3841 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     3901 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     3961 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     4021 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4081 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4141 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4201 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4261 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4321 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4381 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4441 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     4501 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4561 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     4621 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     4681 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     4741 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     4801 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     4861 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     4921 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     4981 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5041 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5101 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5161 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5221 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5281 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     5341 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     5401 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     5461 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     5521 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     5581 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     5641 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     5701 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     5761 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     5821 gcgaaat
//
