LOCUS Exported 6201 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Allows the integration of Myc ArC Long in the genome and Tet-inducible expression.. ACCESSION . VERSION . KEYWORDS pcDNA5 FRT TO Myc ArC Long WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6201) AUTHORS Girdler F, Gascoigne KE, Eyers PA, Hartmuth S, Crafter C, Foote KM, Keen NJ, Taylor SS TITLE Validating Aurora B as an anti-cancer drug target. JOURNAL J Cell Sci. 2006 Sep 1;119(Pt 17):3664-75. Epub 2006 Aug 15. PUBMED 16912073 REFERENCE 2 (bases 1 to 6201) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6201 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 820..838 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 869..893 /label=LNCX /note="Human CMV promoter, forward primer" CDS 1154..1183 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1820..1831 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2153..2170) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2159..2383 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2516..2535) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2667..2714 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2722..3744 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 3874..3995 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3911..3930) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3965..3984 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4044..4060) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4044..4060) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4057..4079) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4068..4084 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4092..4122) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4137..4158 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4275..4292) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4446..5034) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4526..4545) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5205..6065) /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 5828..5847 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6066..6170) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtaccc cggggaattc ggcagatccc gaggtccgac 1021 agcgcccggc ccagatcccc acgcctgcca ggagcaagcc gagagccagc cggccggcgc 1081 actccgactc cgagcagtct ctgtccttcg acccgagccc cgcgcccttt ccgggacccc 1141 tgccccgcgg atggagcaga agctcatctc agaagaagac ctcgagggat ccagctcccc 1201 cagagctgtg gtgcagctgg gcaaagctca acctgcaggc gaagagttgg ctacagcaaa 1261 ccaaacagcc cagcagccca gcagcccagc catgcggcgc ctcacagtcg atgactttga 1321 aatcgggcgt cccctgggca aggggaaatt tgggaatgtg tacctggctc ggctcaagga 1381 aagccatttc attgtggccc tgaaggttct cttcaagtcg cagatagaga aggaaggact 1441 ggagcaccag ctgcgccggg aaattgagat ccaggctcat ctacaacacc ccaatatcct 1501 gcgcctgtat aactatttcc atgatgcacg ccgggtgtac ctgattctgg aatatgctcc 1561 aaggggtgag ctctacaagg agctgcagaa aagcgagaaa ttagatgaac agcgcacagc 1621 cacgataata gaggagttgg cagatgccct gacctactgc catgacaaga aagtgattca 1681 cagagatatt aagccagaga acctgctgct ggggttcagg ggtgaggtga agattgcaga 1741 ttttggctgg tctgtgcaca ccccctccct gaggaggaag acaatgtgtg ggacactgga 1801 ctacttgccg ccagaaatga ttgaggggag aacatatgat gaaaaggtgg atttgtggtg 1861 cattggagtg ctctgctatg agctgctggt gggatatcca ccctttgaga gcgcctccca 1921 cagtgagact tacagacgca tcctcaaggt agatgtgagg tttccactat caatgcctct 1981 gggggcccgg gacttgattt ccaggcttct cagataccag cccttggaga gactgcccct 2041 ggcccagatc ctgaagcacc cctgggttca ggcccactcc cgaagggtgc tgcctccctg 2101 tgctcagatg gcttcctgag cggccgcggg cccgtttaaa cccgctgatc agcctcgact 2161 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 2221 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 2281 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 2341 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 2401 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2461 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2521 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2581 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 2641 gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa 2701 agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt 2761 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct 2821 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc 2881 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt 2941 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt 3001 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg 3061 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc 3121 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt 3181 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc 3241 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc 3301 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc 3361 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc 3421 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg 3481 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc 3541 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac 3601 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg 3661 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc 3721 actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc 3781 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 3841 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 3901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 3961 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 4021 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 4081 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 4141 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 4201 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 4261 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 4321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 4381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 4441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 4501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 4561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 4621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 4681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 4741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 4801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 4861 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 4921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 4981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 5041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 5101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 5161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 5221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 5281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 5341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 5401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 5461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 5521 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 5581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 5641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 5701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 5761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 5821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 5881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 5941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 6001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 6061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 6121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 6181 cgaaaagtgc cacctgacgt c //