LOCUS Exported 6124 bp ds-DNA circular SYN 17-JUL-2006 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR221-P3P2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6124) AUTHORS Invitrogen TITLE Direct Submission JOURNAL Exported Jan 17, 2019 from SnapGene Viewer 4.2.11 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6124 /organism="synthetic DNA construct" /note="/dnas_title=KA901_HA_PolyA_EF1a_GFP_TETa_PolyA" misc_feature complement(268..295) /label=rrnB T2 transcription termination sequence /note="rrnB T2 transcription termination sequence" misc_feature complement(427..470) /label=rrnB T1 transcription termination sequence /note="rrnB T1 transcription termination sequence" primer_bind 517..533 /label=M13 Forward (-40) primer binding site /note="M13 Forward (-40) primer binding site" primer_bind 537..552 /label=M13 Forward (-20) primer binding site /note="M13 Forward (-20) primer binding site" misc_recomb 570..665 /label=attL3 /note="attL3 recombination site" /note="recognized as attL3" misc_feature 686..715 /label=HA epitope tag /note="HA epitope tag" polyA_signal 733..932 /label=SV40 polyA /note="SV40 polyA" promoter 939..2122 /note="EF1a promoter" promoter 2139..2158 /note="T7 promoter" gene 2202..2902 /label=eGFP /note="eGFP" misc_feature 2919..2927 /label=GSG linker /note="GSG linker" misc_feature 2928..2984 /label=P2A self-cleaving peptide /note="P2A self-cleaving peptide" misc_feature 2979..2984 /label=G^P cleavage site /note="G^P cleavage site" gene 2994..3740 /label=Tet-On Advanced /note="Tet-On Advanced" polyA_signal 3763..4205 /label=SV40 polyA /note="SV40 polyA" misc_recomb complement(4236..4334) /label=attL2 /note="attL2 recombination site" /note="recognized as attL2" misc_feature join(4246..6124,1..658) /label=nonstandard type: vector /note="/source=pDONR221-P3P2" /note="/type=Donor Vector" primer_bind complement(4389..4405) /label=M13 Reverse primer binding site /note="M13 Reverse primer binding site" gene 4518..5327 /label=Kanamycin resistance gene /note="Kanamycin resistance gene" rep_origin 5448..6121 /label=f1 origin /note="f1 origin" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa 661 agttgagatc aacaaagctg gatcctaccc atacgatgtg ccagattacg cttgatttaa 721 atagataact gactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc 781 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 841 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 901 cacaaataaa gcattttttt cactgcctcg agtctagacg tgaggctccg gtgcccgtca 961 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg 1021 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct 1081 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt 1141 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg 1201 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca 1261 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg 1321 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg 1381 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc 1441 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa 1501 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc 1561 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat 1621 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg 1681 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag 1741 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga 1801 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc 1861 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg 1921 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga 1981 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc 2041 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt 2101 tcttccattt caggtgtcgt gaggaattag cttggtacta atacgactca ctatagggag 2161 acccaagctg gctaggtgaa ttcgactggc gcgccatggt gagcaagggc gaggagctgt 2221 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 2281 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct 2341 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg 2401 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 2461 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 2521 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 2581 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 2641 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc 2701 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca 2761 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga 2821 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg 2881 ggatcactct cggcatggac gagctgtaca agcgcgccgg atcaggagct actaatttct 2941 ctctgttaaa gcaggcagga gatgtggagg agaatccagg tcctcaattc accatgtcca 3001 gactggacaa gagcaaagtc ataaacggcg ctctggaatt actcaatgga gtcggtatcg 3061 aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact 3121 ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc 3181 atcataccca cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg 3241 ccaagtcatt ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca 3301 cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc 3361 aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg 3421 gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca 3481 ccgattctat gcccccactt ctgagacaag caattgagct gttcgaccgg cagggagccg 3541 aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt 3601 gcgaaagcgg cgggccggcc gacgcccttg acgattttga cttagacatg ctcccagccg 3661 atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc 3721 ttgacatgct ccccgggtaa ctaagtaaga atccagacat gataagatac attgatgagt 3781 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 3841 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca 3901 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc 3961 tctacaaatg tggtatggct gattatgatc ctgcaagcct cgtcgtcctg gccggaccac 4021 gctatctgtg caaggtcccc ggccccggac gcgcgctcca tgagcagagc gcccgccgcc 4081 gaggcgaaga ctcgggcggc gccctgcccg tcccaccagg tcaacaggcg gtaaccggcc 4141 tcttcatcgg gaatgcgcgc gaccttcagc atcgccggca tgtccccctg gcggacggga 4201 agtatccagc tcgaccaggc ttgtcgactc gaagaaccca gctttcttgt acaaagttgg 4261 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa 4321 taaaatcatt atttgccatc cagctgcagg gcggccgcga tatcccctat agtgagtcgt 4381 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt 4441 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca 4501 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt 4561 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag 4621 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg 4681 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg 4741 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac 4801 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag 4861 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt 4921 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga 4981 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac 5041 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg 5101 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg 5161 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg 5221 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg 5281 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta 5341 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac 5401 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga 5461 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 5521 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 5581 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 5641 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 5701 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 5761 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 5821 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 5881 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 5941 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 6001 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 6061 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 6121 tgtt //