LOCUS       sequence-72943-e        3964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector: Position 3, FRT site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3964)
  AUTHORS   Kirchmaier S, Lust K, Wittbrodt J
  TITLE     Golden GATEway cloning--a combinatorial approach to generate fusion 
            and recombination constructs.
  JOURNAL   PLoS One. 2013 Oct 7;8(10):e76117. doi: 
            10.1371/journal.pone.0076117.
  PUBMED    24116091
REFERENCE   2  (bases 1 to 3964)
  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: "PLoS One.";
            date: "2013-10-7"; volume: "8(10):e76117. doi"; pages: " 
            10.1371/journal.pone.0076117"
COMMENT     Sequence Label: pGGEV_3_FRT_+1_OA-
FEATURES             Location/Qualifiers
     source          1..3964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        971..989
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     971..988
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(1008..1041)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     intron          1112..1684
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    1757..1891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1781..1800)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1835..1854
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2043..2060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2214..2802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(2973..3764,3765..3833))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3596..3615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3834..3938)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctacgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagggc gcctccccca
      541 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
      601 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
      661 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
      721 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
      781 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
      841 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
      901 tgggaggtct atataagcaa tgctcgttta gggaaccgcc attctgcctg gggacgtcgg
      961 agcaagcttg atttaggtga cactatagag gtctcaacct ctggatcgaa gttcctatac
     1021 tttctagaga ataggaactt cgtaccacta tgagaccctc atcctcccgg gtaactaagt
     1081 aaggatctcg accgatcctg agaacttcag ggtgagtttg gggacccttg attgttcttt
     1141 ctttttcgct attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt
     1201 tagaatggga agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca
     1261 ctttctactc tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt
     1321 ttcgttaaac tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc
     1381 agattgtaag tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg
     1441 tacttcagca cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct
     1501 gcatataaat tctggctggc gtggaaatat tcttattggt agaaacaact acaccctggt
     1561 catcatcctg cctttctctt tatggttaca atgatataca ctgtttgaga tgaggataaa
     1621 atactctgag tccaaaccgg gcccctctgc taaccatgtt catgccttct tctctttcct
     1681 acagctcctg ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattaat
     1741 tcctcgacgg atcgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     1801 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1861 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1921 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgctctag aactagtctg
     1981 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2221 aggctcggcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2281 ccgacaggac tataaagata ccaggcgttc ccccctggaa gctccctcgt gcgctctcct
     2341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2401 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2641 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2881 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactgcccgt cgtgtagata
     3061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgggaccca
     3121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3361 gttacatgat cccccatgtt gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3421 gtcagaagta agttggccgc agtgttatca ctcatgctta tggcagcact gcataattct
     3481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3961 cctg
//
