LOCUS       sequence-10127-e        3051 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 3051)
  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 3051)
  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: pEB1030
FEATURES             Location/Qualifiers
     source          1..3051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             559..582
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(606..622)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..623)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(665..684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     784..806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(844..862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        930..1034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1035..1103,1104..1895)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1253..1272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2066..2654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2555..2574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2808..2825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2942..2963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2978..2983,2984..3001,3002..3008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3016..3032
                     /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)."
     primer_bind     3021..3043
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3040..3051,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3040..3051,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttagccgcc agcgaggcca cgggcggcct ggatcgcgaa
       61 cgcatcgact tgttgtggaa aatcgctcgc gccggcgccc gttccgcagt gggcaccgag
      121 gcgcgtcgcc agttccgcta cgacggcgac atgaatatcg gcgtgatcac cgatttcgag
      181 ctggaagtgc gcaatgcgct gaacaggcgg gcgcacgccg tcggcgcgca ggacgtggtc
      241 cagcatggca ctgagcagaa caatcctttc ccggaggcag atgagaagat tttcgtcgta
      301 tcggccaccg gtgaaagcca gatgctcacg cgcgggcaac tgaaggaata cattggccag
      361 cagcgcggcg agggctatgt cttctacgag aaccgtgcat acggcgtggc ggggaaaagc
      421 ctgttcgacg atgggctggg agccgcgccc ggcgtgccga gcggacgttc gaagttctcg
      481 ccggatgtac tggaaacggt gccggcgtca cccggattgc ggcggccgtc gctgggcgca
      541 gtggaacgcc actgcaggga ttataaagat gacgatgaca agctagaatt cgatatcaag
      601 cttatcgata ccgtcgacct cgagcaattc atcgatataa ctaagtaata tggtgcactc
      661 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
      721 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
      781 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
      841 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
      901 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      961 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1021 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1081 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1141 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1201 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1261 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1321 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1381 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1441 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1501 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1561 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1621 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1681 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1741 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1801 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1861 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1921 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1981 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2041 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2641 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2701 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2761 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2821 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2881 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2941 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     3001 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc t
//
