LOCUS       sequence-270218-        6663 bp    DNA     circular SYN 15-APR-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 Apr 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 – SacII – SpeI(NheI) - ANXA2 (29-339)
FEATURES             Location/Qualifiers
     source          1..6663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             52..72
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1084..1102)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1098..1145
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1320..1325,1326..1342,1343..1348))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1375..1393)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1461..1565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1566..1634,1635..2426)
                     /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(1784..1803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2597..3185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3086..3105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3339..3356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3371..3510
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3436..3455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3596..3618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3612..3803)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6158..6177
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6197..6274)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6480..6499
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6583..6602
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6583..6601
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6602..6626
                     /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             6641..6663
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tataccatgg gccatcacca tcaccatcac gactacgaca tcccgactac cgaaaacctg
       61 tacttccagg gatcgcatat gccgcggact agtgcctata ctaactttga tgctgagcgg
      121 gatgctttga acattgaaac agccatcaag accaaaggtg tggatgaggt caccattgtc
      181 aacattttga ccaaccgcag caatgaacag agacaggata ttgccttcgc ctaccagaga
      241 aggaccaaaa aggaacttgc atcagcactg aagtcagcct tatctggcca cctggagacg
      301 gtgattttgg gcctattgaa gacacctgct cagtatgacg cttctgagct aaaggcttcc
      361 atgaaggggc tgggaaccga cgaggactct ctcattgaga tcatctgctc cagaaccaac
      421 caggagctgc aggaaattaa cagagtctac aaggaaatgt acaagactga tctggagaag
      481 gacattattt cggacacatc tggtgacttc cgcaagctga tggttgccct ggcaaagggt
      541 agaagagcag aggatggctc tgtcattgat tatgaactga ttgaccaaga tgctcgggat
      601 ctctatgacg ctggagtgaa gaggaaagga actgatgttc ccaagtggat cagcatcatg
      661 accgagcgga gcgtgcccca cctccagaaa gtatttgata ggtacaagag ttacagccct
      721 tatgacatgt tggaaagcat caggaaagag gttaaaggag acctggaaaa tgctttcctg
      781 aacctggttc agtgcattca gaacaagccc ctgtattttg ctgatcggct gtatgactcc
      841 atgaagggca aggggacgcg agataaggtc ctgatcagaa tcatggtctc ccgcagtgaa
      901 gtggacatgt tgaaaattag gtctgagttc aagagaaagt acggcaagtc cctgtactat
      961 tatatccagc aagacactaa gggcgactac cagaaagcgc tgctgtacct gtgtggtgga
     1021 gatgactaac tcgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     1081 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1141 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
     1201 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     1261 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     1321 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
     1381 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     1441 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1561 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1801 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2101 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2281 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2401 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     2761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3301 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3361 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3421 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3481 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3541 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3601 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     3661 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     3721 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     3781 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     3841 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     3901 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     3961 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4021 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4081 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     4141 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4201 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4261 cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg
     4321 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
     4381 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
     4441 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     4501 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     4561 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     4621 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     4681 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     4741 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     4801 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4861 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4921 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4981 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     5041 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     5101 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa ctgtcgtgcc agctgcatta
     5161 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     5221 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     5281 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     5341 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     5401 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     5461 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     5521 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     5581 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     5641 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     5701 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     5761 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     5821 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     5881 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5941 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     6001 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     6061 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     6121 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     6181 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     6241 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     6301 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     6361 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     6421 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     6481 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     6541 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
     6601 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
     6661 aga
//
