LOCUS Exported 6807 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-FRET-ORF1-C3-mCerulean_ORF2-TEV-mAmber SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6807) AUTHORS Self JOURNAL Unpublished. REFERENCE 2 (bases 1 to 6807) 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..6807 /organism="synthetic DNA construct" /mol_type="other DNA" source 557..562 /organism="synthetic DNA construct" /mol_type="other DNA" source 563..1282 /organism="synthetic DNA construct" /mol_type="other DNA" source 1908..2733 /organism="synthetic DNA construct" /mol_type="other DNA" source 2014..2730 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 270..543 /note="TK poly(A) signal" /note="TK polyadenylation signal" CDS complement(563..1282) /codon_start=1 /product="Cerulean" /note="Cerulean" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS complement(1283..1306) /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /note="HRV 3C site" /translation="LEVLFQGP" misc_feature 1307..1371 /note="MCS" misc_feature 1374..1901 /note="BI Promoter" /note="bidirectional promoter from BI16 vector (Sammarco and Grabczyk, PMID: 16212928)" protein_bind 1379..1397 /bound_moiety="Tet repressor" /note="tet operator" protein_bind 1400..1418 /bound_moiety="Tet repressor" /note="tet operator" protein_bind 1857..1875 /bound_moiety="Tet repressor" /note="tet operator" protein_bind 1878..1896 /bound_moiety="Tet repressor" /note="tet operator" misc_feature 1926..1946 /note="TEV-L" /note="enocdes peptide cleaved by TEV protease" misc_feature 1987..2007 /note="TEV-R" /note="enocdes peptide cleaved by TEV protease" CDS 2014..2733 /codon_start=1 /note="mAmber" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGCGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 2765..2989 /note="BGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" protein_bind 3273..3320 /bound_moiety="FLP recombinase" /note="FRT" CDS 3328..4350 /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 4480..4610 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin complement(4993..5666) /direction=LEFT /note="pUC origin" /note="pUC origin color: #66cc99; direction: LEFT" CDS complement(5811..6671) /codon_start=1 /product="beta-lactamase" /note="AmpR" /note="Ampicillin color: #ff9966" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6672..6770) /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 ccttacttgt acagctcgtc catgccgaga gtgatcccgg 601 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca 661 gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg 721 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga 781 tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtcgc 841 tgatggcgtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc 901 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct 961 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg 1021 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc 1081 cccaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga 1141 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga 1201 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca 1261 gctcctcgcc cttgctcacc atgggcccct ggaacagaac ttccagacgc gtttcagagc 1321 ggccgcactg atcttaaggg tactagcgct gatcggcaat tgcgaggtcc taggacgatc 1381 tctatcactg atagggagat ctctatcact gatagggaga gctctgctta tatagacctc 1441 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg 1501 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact 1561 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat 1621 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa 1681 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg 1741 cgtacttggc atatgataca cttgatgtac tctagaaaaa tgtcgtaaca actccgcccc 1801 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctccc 1861 tatcagtgat agagatctcc ctatcagtga tagagatcgt caagcttggt accgagctcg 1921 gatccgaaaa cctgtacttc caaggaaccg gtggtcacct gtacaatcga tcctgcagga 1981 gctagcgaga atttgtattt tcagggtgat atcatggtga gcaagggcga ggagctgttc 2041 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 2101 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctcatctgc 2161 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctcgg ctgcggcctg 2221 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 2281 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 2341 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 2401 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 2461 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc 2521 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc 2581 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc 2641 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 2701 atcactctcg gcatggacga gctgtacaag taactcgagg tttaaacccg ctgatcagcc 2761 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2821 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2881 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2941 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 3001 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 3061 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3121 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3181 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3241 aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct 3301 ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag 3361 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa 3421 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc 3481 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc 3541 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc 3601 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg 3661 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg 3721 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc 3781 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg 3841 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg 3901 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca 3961 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc 4021 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg 4081 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac 4141 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga 4201 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga 4261 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc 4321 cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga ttccaccgcc 4381 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 4441 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 4501 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4561 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 4621 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 4861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 4921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 5221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 5461 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 5761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 5821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 5881 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 5941 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 6001 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 6061 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 6121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 6181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 6241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 6301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 6361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 6421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 6481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 6541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 6601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 6661 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 6721 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 6781 tttccccgaa aagtgccacc tgacgtc //