LOCUS Exported 5483 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 5483) 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..5483 /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 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 653..657 /label=MCS misc_feature 658..678 /label=(965)_pAID_Linker_5'_BamHI_SacI CDS 663..885 /codon_start=1 /label=Linker-mAID-Clover /translation="GSGAGAKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKS SGGPEAAAFVKVSMDGAPYLRKIDLRMYK" CDS 663..680 /codon_start=1 /label=Linker /note="note=""direct strand translation""translation=""GAGA""" /translation="GSGAGA" CDS 681..884 /codon_start=1 /label=mAID /note="note=""direct strand translation""translation=""KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNV MVSCQKSSGGPEAAAFVKVSMDGAPYLRKIDLRMYK""" /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA AAFVKVSMDGAPYLRKIDLRMYK" primer_bind 768..787 /label=(811) mAID seq 1 CDS 891..1406 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW RLCERILA" primer_bind complement(1379..1403) /label=(1843) Nluc-NarI polyA_signal 1537..1658 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(1689..1722) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 1689..1722 /label=LoxP-1 /note="note=""direct strand translation"" translation=""ITSYNVCYTKLX""" promoter 1783..2282 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind 2113..2134 /label=(1800) PGK promoter seq 2 primer_bind 2288..2331 /label=(1782) BSR FOR CDS 2312..2710 /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(2686..2728) /label=(1783) BSR REV polyA_signal 2734..3189 /label=mouse PGK terminator protein_bind complement(3213..3246) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3213..3246 /label=LoxP-2 primer_bind complement(3257..3282) /label=(1007)_Marker_3'_BamHI_KpnI promoter complement(3295..3313) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3334..3350) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3358..3374 /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(3382..3412) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3427..3448 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3736..4324) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4495..5355) /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(5356..5460) /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 tggagctccc 661 cgggatccgg tgcaggcgcc aaggagaaga gtgcttgtcc taaagatcca gccaaacctc 721 cggccaaggc acaagttgtg ggatggccac cggtgagatc ataccggaag aacgtgatgg 781 tttcctgcca aaaatcaagc ggtggcccgg aggcggcggc gttcgtgaag gtatcaatgg 841 acggagcacc gtacttgagg aaaatcgatt tgaggatgta taaagctagc atggtcttca 901 cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc 961 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc 1021 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt 1081 atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc 1141 ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg 1201 ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg 1261 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc 1321 tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc 1381 ggctgtgcga acgcattctg gcgtaagcgg ccgctcgagt ctagataact gatcataatc 1441 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 1501 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 1561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 1621 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgatca tattcaataa 1681 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtctg aagaggagtt 1741 tacgtccagc caagcttagg atctcgacct cgaaattcta ccgggtaggg gaggcgcttt 1801 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa 1861 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg actccgttct 1921 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc 1981 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc 2041 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc 2101 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc 2161 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct 2221 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt 2281 cgacctgcat ccatctagat ctcgagccac catggccaag cctttgtctc aagaagaatc 2341 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag 2401 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca 2461 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc 2521 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg 2581 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga 2641 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg 2701 ggagggctaa tcgatgatat cagatccccg ggatgcagaa attgatgatc tattaaacaa 2761 taaagatgtc cactaaaatg gaagtttttc ctgtcatact ttgttaagaa gggtgagaac 2821 agagtaccta cattttgaat ggaaggattg gagctacggg ggtgggggtg gggtgggatt 2881 agataaatgc ctgctcttta ctgaaggctc tttactattg ctttatgata atgtttcata 2941 gttggatatc ataatttaaa caagcaaaac caaattaagg gccagctcat tcctcccact 3001 catgatctat agatctatag atctctcgtg ggatcattgt ttttctcttg attcccactt 3061 tgtggttcta agtactgtgg tttccaaatg tgtcagtttc atagcctgaa gaacgagatc 3121 agcagcctct gttccacata cacttcattc tcagtattgt tttgccaagt tctaattcca 3181 tcagaagctg gtcgagatcc ggaaccctta atataacttc gtataatgta tgctatacga 3241 agttattagg tccctcgaag aggttcacta ggatccggta cccagctttt gttcccttta 3301 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 3361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 3421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 3481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 3541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 3601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 3661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 4141 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct 4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 4381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 4441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 4501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 4561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 4621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 4681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 4741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 4801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 4861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 4921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 4981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 5041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 5101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 5161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 5221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 5281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 5341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 5401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 5461 cacatttccc cgaaaagtgc cac //