LOCUS Exported 2997 bp ds-DNA circular SYN 21-JUL-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTAT-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2997) AUTHORS Nagahara H, Vocero-Akbani AM, Snyder EL, Ho A, Latham DG, Lissy NA, Becker-Hapak M, Ezhevsky SA, Dowdy SF TITLE Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration. JOURNAL Nat Med. 1998 Dec;4(12):1449-52. PUBMED 9846587 REFERENCE 2 (bases 1 to 2997) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2997 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 51..73 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 93..110 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 114..146 /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 150..173 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 252..278 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(379..397) /label=T7 Term /note="T7 terminator, reverse primer" terminator 393..440 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 537..992 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(624..643) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 834..855 /label=F1ori-F /note="F1 origin, forward primer" promoter 1018..1122 /gene="bla" /label=AmpR promoter CDS 1123..1983 /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(1341..1360) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2154..2742 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2643..2662 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2896..2913 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac 61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta 121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatcgat 181 ggggatccaa gcttggctac ggccgcaaga aacgccgcca gcgccgccgc ggtggatcca 241 ccatgtccgg ctatccatat gacgtcccag actatgctgg ctccatggcc ggtaccggtc 301 tcgaggtgca tgcggtgaat tcgaagcttg atccggctgc taacaaagcc cgaaaggaag 361 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 421 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctggc gtaatagcga 481 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc 541 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 601 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 661 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 721 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 781 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 841 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 901 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 961 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc 1021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 1081 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 1141 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 1201 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 1261 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 1321 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 1381 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 1441 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 1501 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 1561 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 1621 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 1681 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 1741 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 1801 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 1861 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 1921 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 1981 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 2041 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 2101 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 2161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 2221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 2281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 2341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 2401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 2461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 2521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 2581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 2641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 2701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 2761 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 2821 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 2881 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 2941 ccgcctctcc ccgcgcgttg gccgattcat taatgcagga tctcgatccc gcgaaat //