LOCUS       sequence-270221-        6663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ANXA2 chimeras in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6663)
  AUTHORS   Ecsedi P, Gogl G, Hof H, Kiss B, Harmat V, Nyitray L
  TITLE     Structure Determination of the Transactivation Domain of p53 in 
            Complex with S100A4 Using Annexin A2 as a Crystallization Chaperone.
  JOURNAL   Structure. 2020 Aug 4;28(8):943-953.e4. doi: 
            10.1016/j.str.2020.05.001. Epub 2020 May 21.
  PUBMED    32442400
REFERENCE   2  (bases 1 to 6663)
  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"; doi: 
            "10.1016/j.str.2020.05.001"; journalName: "Structure"; date: 
            "2020-08-4- 4"; volume: "28"; issue: "8"; pages: "943-953.e4"
COMMENT     Sequence Label: NdeI – SpeI(NheI) – BamHI/BglII - ANXA2 (29-339)
FEATURES             Location/Qualifiers
     source          1..6663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             36..53
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             75..95
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1107..1125)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1121..1168
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1343..1348,1349..1365,1366..1371))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1398..1416)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1484..1588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1589..1657,1658..2449)
                     /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(1807..1826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2620..3208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3109..3128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3362..3379
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3394..3533
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3459..3478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3619..3641
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3635..3826)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6181..6200
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6220..6297)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6503..6522
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6606..6625
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6606..6624
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6625..6649
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agatatacca tgggccatca ccatcaccat cacgactacg
       61 acatcccgac taccgaaaac ctgtacttcc agggatcgca tatgactagt ggatccgcct
      121 atactaactt tgatgctgag cgggatgctt tgaacattga aacagccatc aagaccaaag
      181 gtgtggatga ggtcaccatt gtcaacattt tgaccaaccg cagcaatgaa cagagacagg
      241 atattgcctt cgcctaccag agaaggacca aaaaggaact tgcatcagca ctgaagtcag
      301 ccttatctgg ccacctggag acggtgattt tgggcctatt gaagacacct gctcagtatg
      361 acgcttctga gctaaaggct tccatgaagg ggctgggaac cgacgaggac tctctcattg
      421 agatcatctg ctccagaacc aaccaggagc tgcaggaaat taacagagtc tacaaggaaa
      481 tgtacaagac tgatctggag aaggacatta tttcggacac atctggtgac ttccgcaagc
      541 tgatggttgc cctggcaaag ggtagaagag cagaggatgg ctctgtcatt gattatgaac
      601 tgattgacca agatgctcgg gatctctatg acgctggagt gaagaggaaa ggaactgatg
      661 ttcccaagtg gatcagcatc atgaccgagc ggagcgtgcc ccacctccag aaagtatttg
      721 ataggtacaa gagttacagc ccttatgaca tgttggaaag catcaggaaa gaggttaaag
      781 gagacctgga aaatgctttc ctgaacctgg ttcagtgcat tcagaacaag cccctgtatt
      841 ttgctgatcg gctgtatgac tccatgaagg gcaaggggac gcgagataag gtcctgatca
      901 gaatcatggt ctcccgcagt gaagtggaca tgttgaaaat taggtctgag ttcaagagaa
      961 agtacggcaa gtccctgtac tattatatcc agcaagacac taagggcgac taccagaaag
     1021 cgctgctgta cctgtgtggt ggagatgact aactcgagat ccggctgcta acaaagcccg
     1081 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1141 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc
     1201 aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg
     1261 ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa
     1321 tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa
     1381 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     1441 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1501 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1561 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1621 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1681 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1741 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1801 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     1861 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1921 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1981 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2041 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2101 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     2161 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2221 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2281 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2341 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2401 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2461 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2521 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2581 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2641 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2701 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2761 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2821 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2881 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2941 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3001 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3061 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3121 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3181 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3241 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3301 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3361 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3421 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     3481 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3541 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3601 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3661 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3721 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     3781 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     3841 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     3901 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     3961 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4021 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4081 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4141 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4201 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4261 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc
     4321 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
     4381 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
     4441 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
     4501 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
     4561 tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag
     4621 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
     4681 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
     4741 ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc
     4801 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4861 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4921 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4981 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     5041 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     5101 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5161 aaactgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5221 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5281 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5341 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5401 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5461 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5521 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5581 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5641 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5701 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5761 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5821 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5881 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5941 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6001 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6061 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6121 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6181 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6241 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6301 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6361 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6421 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6481 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6541 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     6601 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     6661 aat
//
