LOCUS Exported 4163 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV-myc-LC3 (Atg8) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4163) AUTHORS Lee IH, Cao L, Mostoslavsky R, Lombard DB, Liu J, Bruns NE, Tsokos M, Alt FW, Finkel T TITLE A role for the NAD-dependent deacetylase Sirt1 in the regulation of autophagy. JOURNAL Proc Natl Acad Sci U S A. 2008 Mar 4. 105(9):3374-9. PUBMED 18296641 REFERENCE 2 (bases 1 to 4163) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4163 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 505..529 /label=LNCX /note="Human CMV promoter, forward primer" intron 646..742 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" CDS 872..1246 /codon_start=1 /gene="MAP1LC3B" /product="human LC3 protein" /label=LC3B /note="Atg8 family member found on autophagosomes" /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD EDGFLYMVYASQETFGMKLSV" polyA_signal 1276..1410 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1300..1319) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1354..1373 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1521..1537) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1521..1537) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1534..1556) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1545..1561 /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(1569..1599) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1614..1635 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1752..1769) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1923..2511) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2003..2022) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2682..3542) /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 3305..3324 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3543..3647) /gene="bla" /label=AmpR promoter primer_bind 3715..3733 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3771..3793) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3893..3912 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4106..4128 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4120..4137 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4121..4137 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg 541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct 601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt 661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag 721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg 781 gaattgtacc cgcgggccca ccatggcatc aatgcagaag ctgatctcag aggaggacct 841 gcttatggcc atggaggccc gaattccgac catgccgtcg gagaagacct tcaagcagcg 901 ccgcaccttc gaacaaagag tagaagatgt ccgacttatt cgagagcagc atccaaccaa 961 aatcccggtg ataatagaac gatacaaggg tgagaagcag cttcctgttc tggataaaac 1021 aaagttcctt gtacctgacc atgtcaacat gagtgagctc atcaagataa ttagaaggcg 1081 cttacagctc aatgctaatc aggccttctt cctgttggtg aacggacaca gcatggtcag 1141 cgtctccaca ccaatctcag aggtgtatga gagtgagaaa gatgaagatg gattcctgta 1201 catggtctat gcctcccagg agacgttcgg gatgaaattg tcagtgtaga tctctcgagg 1261 taccgcggcc gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca 1321 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt 1381 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt 1441 caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat 1501 gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 1561 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 1621 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 1681 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 1741 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 1801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 1861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 1921 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 1981 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 2041 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 2101 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 2161 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 2221 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 2281 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 2341 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 2401 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 2461 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 2521 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 2581 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 2641 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 2701 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 2761 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 2821 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 2881 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 2941 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 3001 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 3061 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 3121 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 3181 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 3241 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 3301 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 3361 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 3421 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 3481 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 3541 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 3601 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 3661 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 3721 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 3781 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 3841 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 3901 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg 3961 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 4021 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 4081 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag 4141 ttcgagcttg catgcctgca ggt //