LOCUS Exported 5289 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSET FLIPPi-5u SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5289) AUTHORS Gu H, Lalonde S, Okumoto S, Looger LL, Scharff-Poulsen AM, Grossman AR, Kossmann J, Jakobsen I, Frommer WB TITLE A novel analytical method for in vivo phosphate tracking. JOURNAL FEBS Lett. 2006 Oct 30. 580(25):5885-93. PUBMED 17034793 REFERENCE 2 (bases 1 to 5289) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5289 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 20..39 /label=T7 /note="T7 promoter, forward primer" promoter 20..38 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 70..92 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 112..129 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 133..165 /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 169..192 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 1915..2634 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2690..2708) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2704..2751 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2848..3303 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2935..2954) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3145..3166 /label=F1ori-F /note="F1 origin, forward primer" promoter 3329..3433 /gene="bla" /label=AmpR promoter CDS 3434..4294 /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(3652..3671) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4465..5053 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4954..4973 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5207..5224 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat 121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac 181 gatgacgata aggatccggg ccgcatggtg agcaagggcg aggagctgtt caccggggtg 241 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 301 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 361 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc 421 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 481 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 541 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 601 gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat 661 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc 721 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 781 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc 841 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcggtacc 901 gtaggatttc taacagcgac ctcggctcaa gcccaaaccg tgcaaatctc cggggcgggc 961 gcgacctttg cggctccttt gctgcaacgt tggtttgacg cctacaaccg caccgtagac 1021 cccactgtgc aagtcagcta tcagtctgtc ggtgctggtg ctggcctaga gcaggtgatc 1081 aatggcactg tggacttcgg cgcttccgag gcgcctttct ccggtgctcg cctggagagc 1141 ttccgagcta aatacggcta tgatccccta cagttgcctc tggcgggagg ggccatcgag 1201 tttgcctata acctgcccgg cattgaagac ggagagctca tcctgaagcg gaaaacctac 1261 tgcggcatcg tgaccggcga gatcactcgc tgggacgaca ttcgcatcaa ggccgagaac 1321 ccaggtatag caaacaagct gccacccctg gacatcacct gggtacaccg ctctgatggt 1381 tctgggacta cctttgtgtt caccaaccac atcagaactg tctgccctaa ttggacagcc 1441 ggtgctggta cttctgtcga gtggcctgtt ggtattggag cccaagggaa tgagggcgta 1501 gccgccacca tcaagcagga gccaggggcg attggctacg tgaaccagtc ctatgccaag 1561 ctggaaaaga tggccactgc tcgcttggaa aacaaagcgg gcaacattgt tgagttctcg 1621 actgaggcag ctacctcggc gctggatgct cccattcctg atgactttgc gctgttggtg 1681 cccgaccctg aagggccaaa tgactaccca atcgtgggct tgttctgggt gatgctgtac 1741 cgcgagtatc ccgatcagca gaagctgacc aagctggtgg aggctctgaa gtggacccag 1801 gggccagagg gtcaagccat caccaaggag ctggactaca tccctatgcc tgaggcggtt 1861 atccagcgga tctttgcaga gctggattcc atcaccgtta acggtaccgg tggaatggtg 1921 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 1981 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 2041 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 2101 accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac 2161 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 2221 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 2281 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 2341 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 2401 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 2461 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 2521 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 2581 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaagctt 2641 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 2701 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 2761 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 2821 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 2881 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 2941 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3001 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 3061 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 3121 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 3181 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 3241 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 3301 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 3361 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 3421 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 3481 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 3541 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 3601 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 3661 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 3721 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 3781 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 3841 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 3901 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 3961 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 4021 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 4081 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 4141 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 4201 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 4261 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 4321 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 4381 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 4441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 4501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 4561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 4621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 4681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 4741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 4801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 4861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 4921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 4981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 5041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 5101 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 5161 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 5221 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 5281 ttaatgcag //