LOCUS       sequence-10126-e        3468 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 3468)
  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 3468)
  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: pEB1029
FEATURES             Location/Qualifiers
     source          1..3468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             685..708
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(732..748)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(733..749)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(763..785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1150..1944)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1261..1280)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1871..1890
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      2340..2884
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2785..2804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3001..3018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3359..3380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3395..3400,3401..3418,3419..3425)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3433..3449
                     /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     3438..3460
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3457..3468,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3457..3468,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatgc agcaatcgca tcaggctggt tacgcaaacg ccgccgaccg ggagtctggc
       61 atccccgcag ccgtactcga tggcatcaag gccgtggcga aggaaaaaaa cgccacattg
      121 atgttccgcc tggtcaaccc ccattccacc agcctgattg ccgaaggggt ggccaccaaa
      181 ggattgggcg tgcacgccaa gtcgtccgat tgggggttgc aggcgggcta cattcccgtc
      241 aacccgaatc tttccaaact gttcggccgt gcgcccgagg tgatcgcgcg ggccgacaac
      301 gacgtcaaca gcagcctggc gcatggccat accgcggtcg acctgacgct gtcgaaagag
      361 cggcttgact atctgcggca agcgggcctg gtcaccggca tggccgatgg cgtggtcgcg
      421 agcaaccacg caggctacga gcagttcgag tttcgcgtga aggaaacctc ggacgggcgc
      481 tatgccgtgc agtatcgccg caagggcggc gacgatttcg aggcggtcaa ggtgatcggc
      541 aatgccgccg gtattccact gacggcggat atcgacatgt tcgccattat gccgcatctg
      601 tccaacttcc gcgactcggc gcgcagttcg gtgaccagcg gcgattcggt gaccgattac
      661 ctggcgcgca cgcggcgggc tgcagattat aaagatgacg atgacaagct agaattcgat
      721 atcaagctta tcgataccgt cgacctcgag caattcggcc gtcgttttac aacgtcgtga
      781 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      841 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      901 tggcgaatgg cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc
      961 tgaacaggag ggacaggggg tgggcgaaga actccagcat gagatccccg cgctggagga
     1021 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     1081 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     1141 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     1201 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     1261 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     1321 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     1381 atgggtcacg acgagatcct cgccgtcggg catccgcgcc ttgagcctgg cgaacagttc
     1441 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     1501 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     1561 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     1621 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     1681 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     1741 cgatagccgc gctgcctcgt cttggagttc attcagggca ccggacaggt cggtcttgac
     1801 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     1861 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     1921 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct
     1981 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
     2041 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
     2101 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
     2161 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     2221 tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg
     2281 cttgcggcag cgtgaagctg tgcctagaaa tattttatct gattaataag atgatcttct
     2341 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     2401 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     2461 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     2521 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     2581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     2641 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     2701 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     2761 gagagcgcac gagggagccg ccaggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2821 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     2881 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     2941 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     3001 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     3061 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     3121 catagtaagc cagtatatac actccgctaa gctttaatgc ggtagtttat cacagttaaa
     3181 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
     3241 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     3301 gcgggatctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3361 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3421 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagct
//
