LOCUS       sequence-270216-        6681 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 6681)
  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 6681)
  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 (23-339)
FEATURES             Location/Qualifiers
     source          1..6681
                     /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(1125..1143)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1139..1186
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1361..1366,1367..1383,1384..1389))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1416..1434)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1502..1606
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1607..1675,1676..2467)
                     /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(1825..1844)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2638..3226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3127..3146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3380..3397
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3412..3551
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3477..3496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3637..3659
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3653..3844)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6199..6218
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6238..6315)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6521..6540
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6624..6643
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6624..6642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6643..6667
                     /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 ggatccgctt
      121 atgggtctgt caaagcctat actaactttg atgctgagcg ggatgctttg aacattgaaa
      181 cagccatcaa gaccaaaggt gtggatgagg tcaccattgt caacattttg accaaccgca
      241 gcaatgaaca gagacaggat attgccttcg cctaccagag aaggaccaaa aaggaacttg
      301 catcagcact gaagtcagcc ttatctggcc acctggagac ggtgattttg ggcctattga
      361 agacacctgc tcagtatgac gcttctgagc taaaggcttc catgaagggg ctgggaaccg
      421 acgaggactc tctcattgag atcatctgct ccagaaccaa ccaggagctg caggaaatta
      481 acagagtcta caaggaaatg tacaagactg atctggagaa ggacattatt tcggacacat
      541 ctggtgactt ccgcaagctg atggttgccc tggcaaaggg tagaagagca gaggatggct
      601 ctgtcattga ttatgaactg attgaccaag atgctcggga tctctatgac gctggagtga
      661 agaggaaagg aactgatgtt cccaagtgga tcagcatcat gaccgagcgg agcgtgcccc
      721 acctccagaa agtatttgat aggtacaaga gttacagccc ttatgacatg ttggaaagca
      781 tcaggaaaga ggttaaagga gacctggaaa atgctttcct gaacctggtt cagtgcattc
      841 agaacaagcc cctgtatttt gctgatcggc tgtatgactc catgaagggc aaggggacgc
      901 gagataaggt cctgatcaga atcatggtct cccgcagtga agtggacatg ttgaaaatta
      961 ggtctgagtt caagagaaag tacggcaagt ccctgtacta ttatatccag caagacacta
     1021 agggcgacta ccagaaagcg ctgctgtacc tgtgtggtgg agatgactaa ctcgagatcc
     1081 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1141 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1201 tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca
     1261 gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt
     1321 gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata
     1381 agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     1441 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1501 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1561 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1621 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1681 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1741 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1801 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1861 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1921 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1981 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2041 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2101 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     2161 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2221 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2281 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2341 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2401 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2461 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2521 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2761 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3301 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3361 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3421 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     3481 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3541 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3601 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3661 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3721 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3781 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3841 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     3901 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     3961 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     4021 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     4081 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     4141 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     4201 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     4261 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc
     4321 caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt
     4381 tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
     4441 ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
     4501 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat
     4561 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
     4621 tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
     4681 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     4741 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     4801 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     4861 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     4921 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     4981 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     5041 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5101 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa actgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5221 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     5281 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     5341 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     5401 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     5461 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     5521 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     5581 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     5641 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     5701 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     5761 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     5821 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     5881 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     5941 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     6001 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     6061 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     6121 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     6181 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     6241 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     6301 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     6361 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     6421 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6481 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6541 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     6601 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     6661 caattcccct ctagaaataa t
//
