LOCUS Exported 6800 bp ds-DNA circular SYN 28-MAY-2017 DEFINITION Ligation of inverted Fragment 1 into Fragment 2. ACCESSION BI-16_+_RRP6_WT_ VERSION . KEYWORDS pKK-BI16-ORF1-C3-mRuby3_ORF2-TEV-mClover3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6800) AUTHORS Self JOURNAL Unpublished. REFERENCE 2 (bases 1 to 6800) AUTHORS Lukasz S. Borowski TITLE Direct Submission JOURNAL Exported Sunday, May 28, 2017 from SnapGene 3.3.3 http://www.snapgene.com COMMENT SECID/File created by SciEd Central, Scientific & Educational Software SECNOTES|Vector molecule: Fragment 2 Fragment ends: XmaJI and ClaI Fragment size: 7594 Insert molecule: Fragment 1 Fragment ends: XmaJI and ClaI Fragment size: 1494 ORIGDB|GenBank FEATURES Location/Qualifiers source 1..6800 /organism="synthetic DNA construct" /mol_type="other DNA" source 562..1275 /organism="synthetic DNA construct" /mol_type="other DNA" source 1901..2726 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 270..543 /note="TK poly(A) signal" /note="TK polyadenylation signal" CDS complement(562..1275) /codon_start=1 /note="mRuby3" /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV AKYSNLGGGMDELYK" CDS complement(1276..1299) /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /note="HRV 3C site" /translation="LEVLFQGP" misc_feature 1300..1364 /note="MCS" misc_feature 1367..1894 /note="BI Promoter" /note="bidirectional promoter from BI16 vector (Sammarco and Grabczyk, PMID: 16212928)" protein_bind 1372..1390 /bound_moiety="Tet repressor" /note="tet operator" protein_bind 1393..1411 /bound_moiety="Tet repressor" /note="tet operator" protein_bind 1850..1868 /bound_moiety="Tet repressor" /note="tet operator" protein_bind 1871..1889 /bound_moiety="Tet repressor" /note="tet operator" misc_feature 1919..1939 /note="TEV-L" /note="enocdes peptide cleaved by TEV protease" misc_feature 1980..2000 /note="TEV-R" /note="enocdes peptide cleaved by TEV protease" CDS 2007..2726 /codon_start=1 /note="mClover3" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" polyA_signal 2758..2982 /note="BGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" protein_bind 3266..3313 /bound_moiety="FLP recombinase" /note="FRT" CDS 3321..4343 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /note="Hygromycin B" /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4473..4603 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin complement(4986..5659) /direction=LEFT /note="pUC origin" /note="pUC origin color: #66cc99; direction: LEFT" CDS complement(5804..6664) /codon_start=1 /product="beta-lactamase" /note="AmpR" /note="Ampicillin color: #ff9966" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6665..6763) /gene="bla" /note="Amp promoter" ORIGIN 1 gacggatcgg gagatccgct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 61 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 121 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 181 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 241 tctccgctcc atcgttcaga tccttatcga tgctatggca gggcctgccg ccccgacgtt 301 ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac 361 gcgggcgtat tggccccaat ggggtcacgg tggggtatcg acagagtgcc agccctggga 421 ccgaaccccg cgtttatgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta 481 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc 541 cccatcagcg aaccgcgggc cttacttgta cagctcgtcc atgccaccac caaggttgct 601 gtatttggca actgccactt ctctttgcac tacgtaggtt tcattgtcac tctcctcgat 661 cctttccagg cggtgatcaa cggcatggac accgggcatc ttgatgttcc cgacggtctt 721 ttttgacctg taagttgtca cgaagttgca gtgcagatgg ccaccaccat caactttcag 781 tgcgatgtca gtgtatcctc tcagaccacc atctgctgga tacatcatct ctgtattagg 841 ctcccaaccc ttggtcttct tctgcatcac gggaccattg gagggaaagt ttacccctct 901 gaccttgacg ttgtagacga gctcgccatc ctcaaggctg gtgtcctgcg tgacggtgac 961 gactccacca tcttcgtatc tcgtaactct ttcccaagta aaaccctcag gaaaggactg 1021 tttaaagaaa tcagggatgt cggccgggta cttgataaag gtacggctgc catacatgaa 1081 cgacgtggca agaatgtcaa aggcaaatgg caggggtcct ccctcgatga ctttgatcct 1141 catggtttgc actccctcgt acggtctgcc ttctccttca cctgtgcatt tgaattggtg 1201 gccgttgacc gaaccttcca tgaccacctt catacgcata ttttccttga tcagctcttc 1261 gcccttagac accatgggcc cctggaacag aacttccaga cgcgtttcag agcggccgca 1321 ctgatcttaa gggtactagc gctgatcggc aattgcgagg tcctaggacg atctctatca 1381 ctgataggga gatctctatc actgataggg agagctctgc ttatatagac ctcccaccgt 1441 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc 1501 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat 1561 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg 1621 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg 1681 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt 1741 ggcatatgat acacttgatg tactctagaa aaatgtcgta acaactccgc cccattgacg 1801 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt 1861 gatagagatc tccctatcag tgatagagat cgtcaagctt ggtaccgagc tcggatccga 1921 aaacctgtac ttccaaggaa ccggtggtca cctgtacaat cgatcctgca ggagctagcg 1981 agaatttgta ttttcagggt gatatcatgg tgagcaaggg cgaggagctg ttcaccgggg 2041 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtccgcg 2101 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg 2161 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc gtggcctgct 2221 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 2281 gctacgtcca ggagcgcacc atctctttca aggacgacgg tacctacaag acccgcgccg 2341 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 2401 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cactacgtct 2461 atatcacggc cgacaagcag aagaactgca tcaaggctaa cttcaagatc cgccacaacg 2521 ttgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 2581 gccccgtgct gctgcccgac aaccactacc tgagccatca gtccaagctg agcaaagacc 2641 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggattacac 2701 atggcatgga cgagctgtac aagtaactcg aggtttaaac ccgctgatca gcctcgactg 2761 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 2821 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 2881 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 2941 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa 3001 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 3061 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 3121 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3181 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 3241 attagggtga tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa 3301 gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc 3361 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc 3421 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg 3481 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc 3541 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg 3601 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg 3661 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc 3721 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg 3781 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg 3841 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg 3901 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca 3961 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct 4021 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg 4081 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct 4141 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg 4201 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg 4261 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca 4321 ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct 4381 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4441 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4501 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4561 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4621 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4681 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 4741 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 4801 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 4861 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 4921 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 4981 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5041 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5101 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5161 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5221 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5281 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5341 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5401 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5461 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5521 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5581 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 5641 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 5701 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 5761 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 5821 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 5881 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 5941 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6001 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6061 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6121 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6181 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6241 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6301 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6361 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6421 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6481 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6541 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 6601 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 6661 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 6721 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 6781 gaaaagtgcc acctgacgtc //