LOCUS       sequence-389403-        5911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized bacterial expression plasmid. Expresses S100A6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5911)
  AUTHORS   Shihora A, Elias RD, Hammond JA, Ghirlando R, Deshmukh L
  TITLE     ALS Variants of Annexin A11's Proline-Rich Domain Impair Its 
            S100A6-Mediated Fibril Dissolution.
  JOURNAL   ACS Chem Neurosci. 2023 Jul 11. doi: 10.1021/acschemneuro.3c00169.
  PUBMED    37433222
REFERENCE   2  (bases 1 to 5911)
  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: "ACS Chem 
            Neurosci. 2023 Jul 11. doi: 10.1021/acschemneuro.3c00169."
COMMENT     Sequence Label: S100A6
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..64
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(58..249)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1561..2643)
                     /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     2605..2624
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2644..2721)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2927..2946
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3030..3049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3030..3048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3049..3073
                     /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             3088..3110
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3441..3459)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3455..3502
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3677..3682,3683..3699,3700..3705))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3732..3750)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3818..3922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3923..3991,3992..4783)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4141..4160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4954..5542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5443..5462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5696..5713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5728..5867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5793..5812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
       61 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
      121 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
      181 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
      241 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
      301 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
      361 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
      421 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
      481 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
      541 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
      601 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
      661 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
      721 ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga
      781 tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc
      841 aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg
      901 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
      961 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
     1021 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     1081 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     1141 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
     1201 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     1261 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     1321 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     1381 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     1441 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     1501 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     1561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1621 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     1681 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     1741 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     1801 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     1861 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     1921 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     1981 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     2041 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     2101 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     2161 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     2221 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     2281 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     2341 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     2401 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     2461 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     2521 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     2581 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     2641 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     2701 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     2761 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     2821 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     2881 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     2941 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     3001 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     3061 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     3121 gcgtgcccgc tggatcaagc gattggcctg ctggtggcga tttttcataa atatagcggc
     3181 cgcgaaggcg ataaacatac cctgagcaaa aaagaactga aagaactgat tcagaaagaa
     3241 ctgaccattg gcagcaaact gcaagatgcg gaaattgcgc gcctgatgga agatctggat
     3301 cgcaacaaag atcaagaagt gaactttcaa gaatatgtga cctttctggg cgcgctggcg
     3361 ctgatttata acgaagcgct gaaaggctaa ggatccggct gctaacaaag cccgaaagga
     3421 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     3481 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg
     3541 cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg
     3601 atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac
     3661 tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg agaattcttg
     3721 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3781 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4141 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     4201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4441 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     4501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5101 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5701 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     5761 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5821 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     5881 caacacccgc tgacgcgccc tgacgggctt g
//
