LOCUS Exported 5505 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSET FlipSuc-90u delta1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5505) AUTHORS Lager I, Looger LL, Hilpert M, Lalonde S, Frommer WB TITLE Conversion of a putative Agrobacterium sugar-binding protein into a FRET sensor with high selectivity for sucrose. JOURNAL J Biol Chem. 2006 Oct 13. 281(41):30875-83. PUBMED 16912038 REFERENCE 2 (bases 1 to 5505) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5505 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 19..38 /label=T7 /note="T7 promoter, forward primer" promoter 19..37 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 69..91 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 111..128 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 132..164 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 168..191 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 2130..2849 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2905..2923) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2919..2966 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 3063..3518 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3150..3169) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3360..3381 /label=F1ori-F /note="F1 origin, forward primer" promoter 3544..3648 /gene="bla" /label=AmpR promoter CDS 3649..4509 /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(3867..3886) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4680..5268 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5169..5188 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5422..5439 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta 61 gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc 121 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg 181 atgacgataa ggatccgggc cgcatggtga gcaagggcga ggagctgttc accggggtgg 241 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 301 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 361 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca 421 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 481 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 541 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 601 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata 661 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg 721 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 781 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 841 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcggtaccg 901 gaggcgccgc tgaggcggcc gaactttcca ttgccgcaaa ttcgaccggc aagaacgtcg 961 cgtttttccg cgaacgcatc gccgctttcg agaaagagac cggtcacaag gtcaatctcg 1021 tcaccatgcc gtcttcttcc agcgaacagt tcagccagta ccggctgtgg cttgccgccg 1081 gcaacaagga tgtcgacgtt taccagacgg acgtgatctg ggccccgcag ctagccgaac 1141 aattcgtgga tttgaccgca gcgaccaagg atgtcatcgg ggatcatttc ccttctatcg 1201 tcgcctcaca gacggtggac ggcaagctcg tcgccatgcc gatgttcacc gacgcgccgg 1261 cgctgtttta ccgcaaggac cttctggaaa aatatggcaa gcagccgccg aaaacctgga 1321 aggaactgag cgagaccgcc aaggaggttc aggacaagga acgcgcggcc ggccagaagg 1381 acctctgggg cttcgttttt cagggcagcg cctatgaagg tctgacctgc aacgctctgg 1441 aatggatcgc atccgcgggc ggtggccata tcgttgaaac cgacggcgat atctccatca 1501 ataacgagaa ggcggcagct gcaatcgaaa ccgccaaagg ctgggtcggc accatcgcac 1561 cgcagggcgt tctcgcttac aaggaagagg aagcacgcgg tgtctggcag accggcaatt 1621 ccgtcttcat gcgcaactgg ccctatgcct atgcgcttgg aaatggtgcc gacagcgcga 1681 tcaaggacaa gttcggcgtc acgccgctgc cggcaggtga agaaggcgca gcccccgctt 1741 ccacactcgg cggcgcgaac cttgctgttt cgaaatattc cgacgatcag gaagcggcta 1801 tccagctggt aaaattcctg gcatcaaagg atacccagaa gctgcgcgcg atccagctgt 1861 ccaacatgcc gacgatcgct tcgctttacg acgacaagga tgtcgcggcc gcacagccct 1921 tcatgccgac gtggaagccg atcttcgaga ccgccgttcc gcgtccttcc gcctcggcca 1981 aggtgaaata taacgaggtt tcggccaagt tctggggcgc cgtgcacaac acgctctccg 2041 gcaatggtac ggcggcggaa aatctcgaac ttctcgaggt tgaactgacc gatctcaaag 2101 gcaacggctg gggcgccggt accggtggaa tggtgagcaa gggcgaggag ctgttcaccg 2161 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 2221 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 2281 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt 2341 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 2401 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2461 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 2521 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 2581 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 2641 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 2701 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag 2761 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 2821 ctctcggcat ggacgagctg tacaagtaaa agcttgatcc ggctgctaac aaagcccgaa 2881 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct 2941 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tctggcgtaa 3001 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 3061 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgttggtt cacgtagtgg 3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 3421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 3481 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat 3541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 3601 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 3661 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 3721 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 3781 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 3841 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 3901 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 3961 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 4021 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 4081 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 4141 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 4201 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 4261 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 4321 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 4381 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 4441 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 4501 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 4561 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 4621 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 4681 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 4741 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 4801 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 4861 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 4921 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 4981 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 5041 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 5101 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 5161 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 5221 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 5281 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 5341 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 5401 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 5461 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagg //