LOCUS Exported 3964 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Golden Gate entry vector: Position 3, FRT site. ACCESSION . VERSION . KEYWORDS pGGEV_3_FRT_+1_OA- 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3964 /organism="synthetic DNA construct" /mol_type="other DNA" 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(2973..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 //