LOCUS       sequence-393449-        7240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Annexin A7 fibrillization and lipid binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7240)
  AUTHORS   Yu C, Nelson SL, Meisl G, Ghirlando R, Deshmukh L
  TITLE     Phase Separation and Fibrillization of Human Annexin A7 Are Mediated
            by Its Proline-Rich Domain.
  JOURNAL   Biochemistry. 2023 Oct 3. doi: 10.1021/acs.biochem.3c00349.
  PUBMED    37788367
REFERENCE   2  (bases 1 to 7240)
  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: 
            "Biochemistry. 2023 Oct 3. doi: 10.1021/acs.biochem.3c00349."
COMMENT     Sequence Label: Annexin A7-TEV-TwinStrep
FEATURES             Location/Qualifiers
     source          1..7240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(929..2011)
                     /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     1973..1992
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2012..2089)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2295..2314
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2398..2417
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2398..2416
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2417..2441
                     /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             2456..2478
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3956..3976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3995..4000,4001..4024,4025..4060,4061..4084)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(4138..4156)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4152..4199
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4374..4379,4380..4396,4397..4402))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4429..4447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4515..4619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4620..4688,4689..5480)
                     /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(4838..4857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5651..6239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6140..6159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6393..6410
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6425..6564
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6490..6509)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6650..6672
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6666..6857)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
       61 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc
      121 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca
      181 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt
      241 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
      301 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc
      361 cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag
      421 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
      481 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
      541 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc
      601 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
      661 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
      721 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
      781 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
      841 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
      901 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1021 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     1081 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     1141 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     1201 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     1261 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     1321 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     1381 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     1441 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     1501 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     1561 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     1621 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     1681 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     1741 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     1801 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     1861 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     1921 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     1981 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     2041 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     2101 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     2161 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     2221 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     2281 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     2341 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     2401 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     2461 ttaactttaa gaaggagata tacatatgag ctatcctggt tacccgccta ccggctaccc
     2521 tccgtttccg ggatatccac ccgccggtca agaaagctcg ttcccgccga gcgggcagta
     2581 tccctaccct agtggcttcc ctccgatggg cggcggggcg tatccgcaag tcccgagcag
     2641 tggttacccc ggcgctggcg gatatcctgc gcccggcgga tacccggcgc cgggtggata
     2701 tccgggtgcg ccacagcctg gcggggcacc tagctacccc ggtgtccctc cgggccaagg
     2761 cttcggcgtt ccgcccggtg gcgcgggctt ctccggttat cctcagccac cttcgcagtc
     2821 ttacggcggc ggccccgctc aagtacctct gccggggggc tttccggggg gacagatgcc
     2881 gagtcagtat ccgggcggtc agccgaccta tccgagtcag attaacaccg atagctttag
     2941 cagctatccg gtgtttagcc cggtgagcct ggattatagc agcgaaccgg cgaccgtgac
     3001 ccaagtgacc caaggcacca ttcgcccggc ggcgaacttt gatgcgattc gcgatgcgga
     3061 aattctgcgc aaagctatga agggcttcgg taccgacgaa caagcgattg tggatgtggt
     3121 ggcgaaccgc agcaacgatc agcgtcagaa aattaaagcg gcgtttaaaa cgagctatgg
     3181 caaagatctg attaaagatc tgaaaagcga actgagcggc aacatggaag aactgattct
     3241 ggcgctgttt atgccgccga cctattatga tgcgtggagc ttgcgcaaag cgatgcaagg
     3301 cgccggcacc caagaacgcg tgctgattga aattctgtgc acccgcacca accaagaaat
     3361 tcgcgaaatt gtgcgctgct atcagagcga atttggccgc gatctggaaa aagatattcg
     3421 ctcggacaca agcggccact tcgaacgcct gctggtgagc atgtgccaag gcaaccgcga
     3481 tgaaaatcag agcattaacc atcagatggc gcaagaagat gcgcagcgcc tgtatcaagc
     3541 gggcgaaggc cgcctgggca ccgacgagag ctgttttaac atgattctgg cgacccgcag
     3601 cttcccgcag ctgcgcgcga ccatggaagc gtatagccgc atggcgaacc gcgatctgct
     3661 gagcagcgtg agccgcgaat ttagcgggta cgtcgagagc ggcctgaaaa ccattctgca
     3721 gtgcgcgctg aaccgcccgg cgttttttgc ggaacgcctg tattatgcca tgaagggggc
     3781 tggcacagat gatagcaccc tggtgcgcat tgtggtgacc cgcagcgaaa ttgatctggt
     3841 gcagattaaa cagatgtttg cgcagatgta tcagaaaacc ctgggcacca tgattgcggg
     3901 agacacgagc ggcgattatc gccgcctgct gctggcgatt gtgggcagcg gcagcgaaaa
     3961 cctgtatttt caaggcagcg gcagcggctc gggcagcgcg tggagccatc cgcagtttga
     4021 aaaaggaggc ggaagtggcg gcggtagcgg cggatcagcg tggagccatc cgcagtttga
     4081 aaaataagga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     4141 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     4201 tgaaaggagg aactatatcc ggatatcccg caagaggccc ggcagtaccg gcataaccaa
     4261 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     4321 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     4381 cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac
     4441 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4861 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5161 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5641 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5701 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5761 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5821 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5881 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5941 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6001 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6061 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6361 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6421 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     6481 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     6541 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     6601 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     6661 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     6721 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     6781 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     6841 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     6901 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     6961 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     7021 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     7081 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     7141 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     7201 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
//
