LOCUS Exported 6116 bp ds-DNA circular SYN 24-JUN-2015 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBigT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6116) AUTHORS Addgene TITLE Direct Submission JOURNAL Exported Wednesday, Jun 24, 2015 from SnapGene 2.7.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6116 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 181..197 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." primer_bind 205..221 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 242..260 /note="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 332..556 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" primer_bind 600..616 /note="KS primer" /note="common sequencing primer, one of multiple similar variants" protein_bind 651..684 /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 713..847 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" polyA_signal 967..1101 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" polyA_signal 1221..1355 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" CDS complement(1974..2777) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(2791..3290) /note="PGK promoter" /note="mouse phosphoglycerate kinase 1 promoter" protein_bind 3320..3353 /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(3367..3383) /note="SK primer" /note="common sequencing primer, one of multiple similar variants" promoter complement(3547..3565) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3572..3588) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(3729..4184) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4211..4315 /gene="bla" /note="AmpR promoter" CDS 4316..5176 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 5347..5935 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg 241 aaattaaccc tcactaaagg gaacaaaagc tggtaccgtt ggcgcgccaa ctagaggatc 301 gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc 361 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata 421 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg 481 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg 541 caactagaag gcacagtcga ggctgatcag cgagctcgcg gccgcccgcg ggggcccctc 601 gaggtcgacg gtatcgataa gctagcttgg gctgcaggtc gagggaccta ataacttcgt 661 atagcataca ttatacgaag ttatattaag ggttccggat cagcttgatg gggatccaga 721 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 781 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 841 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga 901 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga 961 gtcgcagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg 1021 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt 1081 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag 1141 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat 1201 tatgatcctc tagagtcgca gatccagaca tgataagata cattgatgag tttggacaaa 1261 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt 1321 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta 1381 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat 1441 gtggtatggc tgattatgat cctctagagt cgcagatcct ctagagtcgc agatctgcaa 1501 gcttctgatg gaattagaac ttggcaaaac aatactgaga atgaagtgta tgtggaacag 1561 aggctgctga tctcgttctt caggctatga aactgacaca tttggaaacc acagtactta 1621 gaaccacaaa gtgggaatca agagaaaaac aatgatccca cgagagatct atagatctat 1681 agatcatgag tgggaggaat gagctggccc ttaatttggt tttgcttgtt taaattatga 1741 tatccaacta tgaaacatta tcataaagca atagtaaaga gccttcagta aagagcaggc 1801 atttatctaa tcccacccca cccccacccc cgtagctcca atccttccat tcaaaatgta 1861 ggtactctgt tctcaccctt cttaacaaag tatgacagga aaaacttcca ttttagtgga 1921 catctttatt gtttaataga tcatcaattt ctgcagactt acagcggatc ccctcagaag 1981 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa 2041 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc 2101 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa 2161 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga 2221 tcatcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc 2281 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct 2341 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc 2401 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac 2461 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca 2521 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc 2581 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc 2641 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag 2701 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt 2761 tcaatggccg atcccatatt ggctgcaggt cgaaaggccc ggagatgagg aagaggagaa 2821 cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg 2881 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag 2941 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac 3001 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc 3061 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg 3121 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt 3181 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc ccagcggggc 3241 tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc ggtagaattc 3301 ctgcaggtcg agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg 3361 gttccggatc cactagttct agagcggcct cgactctacg ataccgtcga tccccactgg 3421 aaagaccgcg aagagtttgt cctcaaccgc gagctgtgga aaaaaaaggg acaggataag 3481 tatgacatca tcaaggaaac cctggactac tgcgccctac agatccctta attaaggcca 3541 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact 3601 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct 3661 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg 3721 gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 3781 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 3841 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 3901 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 3961 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 4021 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 4081 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcagtca cgctgcgcgt 4141 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg 4201 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 4261 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 4321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 4381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 4441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 4501 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 4561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 4621 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 4681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 4741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 4801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 4861 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 4921 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 4981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 5041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 5101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 5161 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 6001 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 6061 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag //