LOCUS Exported 4877 bp ds-DNA circular SYN 11-DEC-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4877) AUTHORS Molecular Function Laboratory TITLE Direct Submission JOURNAL Exported Dec 11, 2018 from SnapGene Viewer 4.2.9 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4877 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 4..459 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 482..501 /label=(852)_LacZ_alpha primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 600..616 /label=(1) M13-20 primer promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 659..676 /codon_start=1 /label=Linker /translation="GSGAGA" CDS 677..721 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 734..799 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 804..813 /codon_start=1 /label=BamHI-Linker-mCherry2 /translation="YK" gene 804..810 /gene="mCherry (variant)" /label=mCherry (variant) polyA_signal 931..1052 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(968..987) /label=(315) SV40 poly A rev misc_feature 1083..1116 /label=LoxP-1 /note="note=""direct strand translation"" translation=""ITSYNVCYTKLX""" protein_bind complement(1083..1116) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 1177..1676 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind 1507..1528 /label=(1800) PGK promoter seq 2 primer_bind 1682..1725 /label=(1782) BSR FOR CDS 1706..2104 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD (Aspergillus terreus cytidine deaminase) /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" primer_bind complement(2080..2122) /label=(1783) BSR REV polyA_signal 2128..2583 /label=mouse PGK terminator misc_feature 2607..2640 /label=LoxP-2 protein_bind complement(2607..2640) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter complement(2689..2707) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2728..2744) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 2752..2768 /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(2776..2806) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2821..2842 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3130..3718) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3889..4749) /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" promoter complement(4750..4854) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg 481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcgg 661 atccggtgca ggcgccaagg aaaccgccgc tgcaaaattc gaacgccagc acatggacag 721 ccgatcccgg gctgactaca aagaccatga cggtgattat aaagatcatg acatcgacta 781 caaggatgac gatgacaagt agtgtacaag taatctagat aactgatcat aatcagccat 841 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 901 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 961 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 1021 tgtggtttgt ccaaactcat caatgtatct taacgcgtcg atcatattca ataaccctta 1081 atataacttc gtataatgta tgctatacga agttattagg tctgaagagg agtttacgtc 1141 cagccaagct taggatctcg acctcgaaat tctaccgggt aggggaggcg cttttcccaa 1201 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc 1261 tctggcctcg cacacattcc acatccaccg gtaggcgcca accgactccg ttctttggtg 1321 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc 1381 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg 1441 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt 1501 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg 1561 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc 1621 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct 1681 gcatccatct agatctcgag ccaccatggc caagcctttg tctcaagaag aatccaccct 1741 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc 1801 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac 1861 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa 1921 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg 1981 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga 2041 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg 2101 ctaatcgatg atatcagatc cccgggatgc agaaattgat gatctattaa acaataaaga 2161 tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga gaacagagta 2221 cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg gattagataa 2281 atgcctgctc tttactgaag gctctttact attgctttat gataatgttt catagttgga 2341 tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc cactcatgat 2401 ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc actttgtggt 2461 tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga gatcagcagc 2521 ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat tccatcagaa 2581 gctggtcgag atccggaacc cttaatataa cttcgtataa tgtatgctat acgaagttat 2641 taggtccctc gaagaggttc actaggatcc ggtacccagc ttttgttccc tttagtgagg 2701 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 2761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 2821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 2881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 2941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 3001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 3061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 3121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 3181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 3241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 3301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 3361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 3421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 3481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 3541 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 3601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 3661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 3721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 3781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 3841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 3901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 3961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 4021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 4081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 4141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 4201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 4261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 4321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 4381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 4441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 4501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 4561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 4621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 4681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 4741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 4801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 4861 tccccgaaaa gtgccac //