LOCUS Exported 7429 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION N-terminal HA-tagged protein expression in mammaiian cells. ACCESSION . VERSION . KEYWORDS HA-Vps34 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7429) AUTHORS Park JM, Jung CH, Seo M, Otto NM, Grunwald D, Kim KH, Moriarity B, Kim YM, Starker C, Nho RS, Voytas D, Kim DH TITLE The ULK1 complex mediates MTORC1 signaling to the autophagy initiation machinery via binding and phosphorylating ATG14. JOURNAL Autophagy. 2016;12(3):547-64. doi: 10.1080/15548627.2016.1140293. PUBMED 27046250 REFERENCE 2 (bases 1 to 7429) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7429 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 14..393 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 394..597 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 548..568 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 594..618 /label=LNCX /note="Human CMV promoter, forward primer" promoter 828..846 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 828..845 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 859..878 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 963..972 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 975..1001 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" regulatory 3729..3738 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 3742..3876 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3779..3798) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3833..3852 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 3945..4302 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3968..3988) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4153..4288 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4215..4234 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4322..4339) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4322..4338) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4331..4353) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4551..5006 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4638..4657) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4848..4869 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5023..5042) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5142..5164 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5202..5220) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5288..5392 /gene="bla" /label=AmpR promoter CDS 5393..6253 /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 complement(5611..5630) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6424..7012 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6913..6932 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7166..7183 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7300..7321 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7336..7366 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7374..7390 /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)." primer_bind 7379..7401 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7398..7414 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7398..7414 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca 61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt 601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga 661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt 721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta 781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac 841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc 901 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc 961 gcgccaccat gggctacccc tatgatgtgc ctgactacgc agatctcaat ggtggtggtg 1021 gtgggtcgac catgggggaa gcagagaagt ttcactacat ctatagttgt gacctggata 1081 tcaacgtcca gcttaagata ggaagcttgg aagggaagag agaacaaaag agttataaag 1141 ctgtcctgga agacccaatg ttgaagttct caggactata tcaagagaca tgctctgatc 1201 tttatgttac ttgtcaagtt tttgcagaag ggaagccttt ggccttgcca gtgagaacat 1261 cctacaaagc atttagtaca agatggaact ggaatgaatg gctgaaacta ccagtaaaat 1321 accctgacct gcccaggaat gcccaagtgg ccctcaccat atgggatgtg tatggtcccg 1381 gaaaagcagt gcctgtagga ggaacaacgg tttcgctctt tggaaaatac ggcatgtttc 1441 gccaagggat gcatgacttg aaagtctggc ctaatgtaga agcagatgga tcagaaccca 1501 caaaaactcc tggcagaaca agtagcactc tctcagaaga tcagatgagc cgtcttgcca 1561 agctcaccaa agctcatcga caaggacaca tggtgaaagt agattggctg gatagattga 1621 catttagaga aatagaaatg ataaatgaga gtgaaaaacg aagttctaat ttcatgtacc 1681 tgatggttga atttcgatgt gtcaagtgtg atgataagga atatggtatt gtttattatg 1741 aaaaggacgg tgatgaatca tctccaattt taacaagttt tgaattagtg aaagttcctg 1801 acccccagat gtctatggag aatttagttg agagcaaaca ccacaagctt gcccggagtt 1861 taagaagtgg accttctgac cacgatctga aacccaatgc tgccacgaga gatcagttaa 1921 atattattgt gagttatcca ccaaccaagc aacttacata tgaagaacaa gatcttgttt 1981 ggaagtttag atattatctt acgaatcaag aaaaagcctt gacaaaattc ttgaaatgtg 2041 ttaattggga tctacctcaa gaggccaaac aggccttgga acttctggga aaatggaagc 2101 cgatggatgt agaggactcc ttggagctgt tatcctctca ttacaccaac ccaactgtga 2161 ggcgttatgc tgttgcccgg ttgcgacagg ccgatgatga ggatttgttg atgtacctat 2221 tacaattggt ccaggctctc aaatatgaaa attttgatga tataaagaat ggattggaac 2281 ctaccaagaa ggatagtcag agttcagtgt cagaaaatgt gtcaaattct ggaataaatt 2341 ctgcagaaat agatagctcc caaattataa ccagccccct tccttcagtc tcttcacctc 2401 ctcctgcatc aaaaacaaaa gaagttccag atggcgaaaa tctggaacaa gatctctgta 2461 ccttcttgat atcgagagcc tgcaaaaact caacactggc taattattta tactggtatg 2521 tgatagtgga atgtgaagat caagatactc agcagagaga tccaaagacc catgagatgt 2581 acttgaacgt aatgagaaga ttcagccaag cattgttgaa gggtgataag tctgtcagag 2641 ttatgcgttc tttgctggct gcacaacaga catttgtaga tcggttggtg catctaatga 2701 aggcagtaca acgcgaaagt ggaaatcgta agaaaaagaa tgagagacta caggcattgc 2761 ttggagataa tgaaaagatg aatttgtcag atgtggaact tatcccgttg cctttagaac 2821 cccaagtgaa aattagagga ataattccgg aaacagctac actgtttaaa agtgccctta 2881 tgcctgcaca gttgtttttt aagacggaag atggaggcaa atatccagtt atatttaagc 2941 atggagatga tttacgtcaa gatcaactta ttcttcaaat catttcactc atggacaagc 3001 tgttacggaa agaaaatctg gacttgaaat tgacacctta taaggtgtta gccaccagta 3061 caaaacatgg cttcatgcag tttatccagt cagttcctgt ggctgaagtt cttgatacag 3121 agggaagcat tcagaacttt tttagaaaat atgcaccaag tgagaatggg ccaaatggga 3181 ttagtgctga ggtcatggac acttacgtta aaagctgtgc tggatattgc gtgatcacct 3241 atatacttgg agttggagac aggcacctgg ataacctttt gctaacaaaa acaggcaaac 3301 tcttccacat agactttgga tatattttgg gtcgggatcc aaagcctctt cctccaccaa 3361 tgaagctgaa taaagaaatg gtagaaggaa tggggggcac acagagtgag cagtaccaag 3421 agttccgtaa acagtgttac acggctttcc tccacctgcg aaggtattct aatctgattt 3481 tgaacttgtt ttccttgatg gttgatgcaa acattccaga tattgcactt gaaccagata 3541 aaactgtgaa aaaggttcag gataaattcc gcttagacct gtcggatgaa gaggctgtgc 3601 attacatgca gagtctgatt gatgagagtg tccatgctct ttttgctgca gtggtggaac 3661 agattcacaa gtttgcccag tactggagaa aatgagcggc cgctaagtaa gtaaggatcc 3721 ccagcttggc cgccatggcc caacttgttt attgcagctt ataatggtta caaataaagc 3781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 3841 tccaaactca tcaatgtatc ttatcatgtc tggatcggga attaattcgg cgcagcacca 3901 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac 3961 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 4021 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 4081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 4141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 4201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 4261 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt aacagcttgg 4321 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 4381 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 4441 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc 4501 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg 4561 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 4621 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 4681 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 4741 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg 4801 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 4861 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt 4921 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 4981 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc 5041 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 5101 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 5161 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt 5221 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 5281 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 5341 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 5401 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 5461 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 5521 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 5581 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 5641 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 5701 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 5761 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 5821 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 5881 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 5941 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 6001 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 6061 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 6121 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 6181 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 6241 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 6301 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 6361 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 6421 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 6481 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 6541 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 6601 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 6661 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 6721 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 6781 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 6841 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 6901 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 6961 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 7021 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 7081 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 7141 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 7201 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 7261 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca 7321 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag 7381 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaattaa //