LOCUS       sequence-328547-        6244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of rat His-tagged Delta-192Annexin A11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6244)
  AUTHORS   Lillebostad PAG, Raasakka A, Hjellbrekke SJ, Patil S, Rostbo T, 
            Hollas H, Sakya SA, Szigetvari PD, Vedeler A, Kursula P
  TITLE     Structure of the ALS Mutation Target Annexin A11 Reveals a 
            Stabilising N-Terminal Segment.
  JOURNAL   Biomolecules. 2020 Apr 24;10(4). pii: biom10040660. doi: 
            10.3390/biom10040660.
  PUBMED    32344647
REFERENCE   2  (bases 1 to 6244)
  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: 
            "Biomolecules."; date: "2020-04-24"; volume: "10(4). pii: 
            biom10040660. doi"; pages: " 10.3390/biom10040660"
COMMENT     Sequence Label: pETM10 with rat Delta-192Annexin A11
FEATURES             Location/Qualifiers
     source          1..6244
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1109..1126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1179..1197)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1193..1240
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1277..1732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1364..1383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1574..1595
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1824..2639)
                     /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(1824..2639)
                     /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     2547..2566
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2631..2650
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2761..3349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3250..3269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3503..3520
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3535..3674
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3600..3619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3760..3782
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3776..3967)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4776..5858)
                     /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     5820..5839
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5859..5936)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6142..6161
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca ccccatgggt
      121 accgatgctt ctggatttga ccccttgcga gatgctgagg tcctgcggaa ggctatgaag
      181 ggcttcggaa cggatgaaca ggccattatc gactgcctag ggagccggtc caacaagcag
      241 cggcagcaga tcctgctgtc cttcaagacg gcctatggca aggatctgat caaagaccta
      301 aagtcagagc tgtcaggaaa cttcgagaag actatcctgg ccctgatgaa gaccccggtc
      361 ctgtttgatg tctatgagat aaaggaagct atcaaggggg ctggtactga tgaagcctgc
      421 ctgattgaga tcctggcttc ccgaagtaac gagcacatcc gggagctaaa cagagcctac
      481 aaaacagagt tcaaaaagac tttggaggag gccattcgaa gtgacacatc aggacacttc
      541 cagagactcc tcatctctct ctctcaggga aaccgtgatg aaagcacaaa cgtggacatg
      601 tcccttgtcc agagagatgt ccaggagctg tacgcagctg gggagaaccg cctgggaaca
      661 gatgaatcca agttcaacgc aattctgtgc tcccggagcc gggcccacct cgtggcagtt
      721 ttcaacgagt atcagaggat gacaggccga gacatcgaga agagcatctg ccgggagatg
      781 tctggggacc tggagcaggg catgctggct gtggtgaaat gcctcaagaa cacccctgcc
      841 ttctttgccg aaaggctcaa caaggctatg aggggagcag gaacaaaaga ccggaccctg
      901 attcgcatca tggtgtctcg aagcgagctt gacctgctgg atatccgagc agagtataag
      961 cggatgtacg gcaagtcact gtaccacgat atcacgggag acacttctgg ggattatcgg
     1021 aagatcctgc tgaagatctg tggtggcaac gactgaggta ccggatccga attcgagctc
     1081 cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc
     1141 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1201 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1261 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1321 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1381 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1441 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1501 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1561 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1621 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1681 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     1741 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1801 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
     1861 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     1921 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     1981 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     2041 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     2101 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     2161 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     2221 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     2281 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     2341 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     2401 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     2461 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     2521 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     2581 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     2641 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
     2701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2881 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3001 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3061 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3421 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3481 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3541 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     3601 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3661 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3721 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3781 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3841 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3901 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3961 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     4021 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     4081 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     4141 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     4201 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4261 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4321 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4381 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     4441 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     4501 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     4561 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     4621 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     4681 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     4741 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4861 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     4921 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     4981 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     5041 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     5101 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     5161 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     5221 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     5281 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     5341 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     5401 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     5461 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     5521 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     5581 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     5641 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     5701 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     5761 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     5821 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     5881 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     5941 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     6001 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     6061 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6121 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6181 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     6241 aaat
//
