LOCUS       sequence-328545-        6820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of rat His-tagged FL Annexin A11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6820)
  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 6820)
  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 FL Annexin A11
FEATURES             Location/Qualifiers
     source          1..6820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1549..1566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1619..1637)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1633..1680
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1717..2172
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1804..1823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2014..2035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2264..3079)
                     /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(2264..3079)
                     /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     2987..3006
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3071..3090
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3201..3789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3690..3709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3943..3960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3975..4114
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4040..4059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4200..4222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4216..4407)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5216..6298)
                     /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     6260..6279
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6299..6376)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6582..6601
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6685..6704
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6685..6703
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6704..6728
                     /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             6743..6765
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6778..6795
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atccaccacc tgctggtggc taccctccag gtgcaccagg tggtgctccc tggggaggtg
       61 ctagctaccc tcctcccagc atgcccccta tcgggctgga taacgtggcc aactatgcag
      121 ggcagttcaa ccaggactac ctctcgggca tggcagccaa catgtctgga acattcggag
      181 gagccaacgt gccaaacatg tatcctgggg ctcctggagg tggctatcct ccagtccctc
      241 ctggtggctt tgggcagccc cctcctgccc agcagcctgt ccctccttat ggaatgtatc
      301 caccgccagg aggaaaccca cctcctggga tgccctcata cccagcatac ccaggggccc
      361 cggtgccagg ccagcctatg ccaccccctg ggcagcagcc cccaggagcc tatcctggac
      421 agcctccaat gacctatcct gggcagtcac caatgccacc ccctgggcag cagccagtgc
      481 caagttaccc aggatactca ggatccagta ccatcacccc ggctgtgcct ccagctcagt
      541 ttggaaacag aggcaccatc accgatgctt ctggatttga ccccttgcga gatgctgagg
      601 tcctgcggaa ggctatgaag ggcttcggaa cggatgaaca ggccattatc gactgcctag
      661 ggagccggtc caacaagcag cggcagcaga tcctgctgtc cttcaagacg gcctatggca
      721 aggatctgat caaagaccta aagtcagagc tgtcaggaaa cttcgagaag actatcctgg
      781 ccctgatgaa gaccccggtc ctgtttgatg tctatgagat aaaggaagct atcaaggggg
      841 ctggtactga tgaagcctgc ctgattgaga tcctggcttc ccgaagtaac gagcacatcc
      901 gggagctaaa cagagcctac aaaacagagt tcaaaaagac tttggaggag gccattcgaa
      961 gtgacacatc aggacacttc cagagactcc tcatctctct ctctcaggga aaccgtgatg
     1021 aaagcacaaa cgtggacatg tcccttgtcc agagagatgt ccaggagctg tacgcagctg
     1081 gggagaaccg cctgggaaca gatgaatcca agttcaacgc aattctgtgc tcccggagcc
     1141 gggcccacct cgtggcagtt ttcaacgagt atcagaggat gacaggccga gacatcgaga
     1201 agagcatctg ccgggagatg tctggggacc tggagcaggg catgctggct gtggtgaaat
     1261 gcctcaagaa cacccctgcc ttctttgccg aaaggctcaa caaggctatg aggggagcag
     1321 gaacaaaaga ccggaccctg attcgcatca tggtgtctcg aagcgagctt gacctgctgg
     1381 atatccgagc agagtataag cggatgtacg gcaagtcact gtaccacgat atcacgggag
     1441 acacttctgg ggattatcgg aagatcctgc tgaagatctg tggtggcaac gactgaggta
     1501 ccggatccga attcgagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac
     1561 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1621 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1681 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     1741 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1801 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1861 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1921 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1981 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2041 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2101 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2161 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2221 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     2281 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2341 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2401 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2461 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     2521 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     2581 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     2641 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2701 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2761 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2821 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2881 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2941 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     3001 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     3061 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     3121 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3181 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3241 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3301 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3361 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3421 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3481 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3541 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3601 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3661 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3721 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3781 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3841 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3901 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3961 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4021 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     4081 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     4141 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4201 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     4261 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     4321 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     4381 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4441 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     4501 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4561 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4621 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4681 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4741 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4801 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4861 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     4921 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     4981 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     5041 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     5101 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5161 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     5221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5281 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     5341 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     5401 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     5461 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     5521 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     5581 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     5641 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     5701 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     5761 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     5821 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     5881 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     5941 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     6001 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     6061 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     6121 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     6181 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     6241 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     6301 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     6361 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     6421 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     6481 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     6541 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     6601 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     6661 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     6721 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac catgaaacat
     6781 caccatcacc atcaccccat gggtatgagc tatccaggct
//
