LOCUS       sequence-10125-e        5061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5061)
  AUTHORS   Battesti A, Bouveret E
  TITLE     Improvement of bacterial two-hybrid vectors for detection of fusion 
            proteins and transfer to pBAD-tandem affinity purification, 
            calmodulin binding peptide, or 6-histidine tag vectors.
  JOURNAL   Proteomics. 2008 Nov . 8(22):4768-71.
  PUBMED    18924111
REFERENCE   2  (bases 1 to 5061)
  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: "Proteomics.
            2008 Nov . 8(22):4768-71."
COMMENT     Sequence Label: pEB587
FEATURES             Location/Qualifiers
     source          1..5061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1350..1523
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1524..1697
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     CDS             1734..1754
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1761..1838
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     primer_bind     complement(1918..1935)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2088..2174
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2266..2293
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2312..2403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2404..2472,2473..3264)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2622..2641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3306..3761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3393..3412)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3872..4460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4361..4380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4614..4631
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4797..4819
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4958..4977
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tggcaggcct tgcgcaacac gatgaagccg tggacaacaa attcaacaaa gaacaacaaa
     1381 acgcgttcta tgagatctta catttaccta acttaaacga agaacaacga aacgccttca
     1441 tccaaagttt aaaagatgac ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc
     1501 taaatgatgc tcaggcgccg aaagtagaca acaaattcaa caaagaacaa caaaacgcgt
     1561 tctatgagat cttacattta cctaacttaa acgaagaaca acgaaacgcc ttcatccaaa
     1621 gtttaaaaga tgacccaagc caaagcgcta accttttagc agaagctaaa aagctaaatg
     1681 gtgctcaggc gccgaaagta gacgcgaatt gtgatatacc tacaactgct tctgaaaatt
     1741 tatattttca aggtgaacta aagagaagat ggaaaaagaa tttcatagcc gtctcagcag
     1801 ccaaccgctt taagaaaatc tcatcctccg gggcacttgt cgagtcgact ctagactcga
     1861 gttcgacctg caggcatgca agcttggctg ttttggcgga tgagagaaga ttttcagcct
     1921 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     1981 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     2041 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     2101 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     2161 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     2221 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     2281 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     2341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2641 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2881 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2941 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3301 gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3361 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3421 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3481 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     3541 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3601 tagtggactc ttgttccaaa cttgaacaac actcaaccct atctcgggct attcttttga
     3661 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3721 atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga
     3781 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3841 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3901 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3961 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4021 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4081 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4141 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4201 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4261 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4321 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4381 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4441 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4501 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4561 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4621 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4681 gtatttcaca ccgcataggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4741 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4801 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcaaggaga
     4861 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     4921 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     4981 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     5041 gctcatgttt gacagcttat c
//
