LOCUS Exported 8679 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses SECFP-hATG16A1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pMRX-IP/SECFP-hATG16A1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8679) AUTHORS Koyama-Honda I, Itakura E, Fujiwara TK, Mizushima N TITLE Temporal analysis of recruitment of mammalian ATG proteins to the autophagosome formation site. JOURNAL Autophagy. 2013 Oct;9(10):1491-9. doi: 10.4161/auto.25529. Epub 2013 Jul 10. PUBMED 23884233 REFERENCE 2 (bases 1 to 8679) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8679 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(529..1128) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(612..632) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 1109..1128 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" misc_feature 1140..1692 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1302..1321) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 1531..1548 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS complement(3598..4317) /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGILTLGMDELYK" primer_bind complement(3643..3664) /label=EGFP-C /note="EGFP, forward primer" primer_bind 3992..4011 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4251..4272 /label=EGFP-N /note="EGFP, reverse primer" misc_feature 4349..4723 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind complement(4395..4411) /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS complement(4733..5149) /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind complement(4749..4765) /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(4781..4803) /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" misc_feature 5214..5571 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" enhancer 5930..6309 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind complement(6411..6429) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6497..6601 /gene="bla" /label=AmpR promoter CDS 6602..7462 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6820..6839) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7626..8214 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8115..8134 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8368..8385 /label=L4440 /note="L4440 vector, forward primer" LTR 8515..305 /note="long terminal repeat from Moloney murine leukemia virus" ORIGIN 1 cttttattga gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt 61 tagttcaaat aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg 121 gttctctaga aactgctgag ggctggaccg catctgggga ccatctgttc ttggccctga 181 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc 241 tgttcttggc cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata 301 ttcagctgtt ccatctgttc ctgaccttga tctgaacttc tctattctca gttatgtatt 361 tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc 421 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatcg tcgagccatg 481 gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc aggcaccggg 541 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt 601 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg 661 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt 721 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg 781 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc 841 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt 901 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac 961 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccgggtcgac 1021 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggcccgggg 1081 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg ttgtggccat 1141 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt 1201 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca 1261 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc 1321 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg 1381 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt 1441 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa 1501 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc 1561 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg 1621 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc 1681 ggccagtaac gttagggggg ggggcggaat ttacgtagcg gccgctcgag ggatcccggg 1741 cccgcggtac cgtcgactgc agaattctca gtactgtgcc cacagcacag ctttgcatcc 1801 tttgtccaca ctgacaacgt gcgagccaga gggcgaccac gccaccgcat tgatggatga 1861 gctgtgctgc tttgaaagaa ccttttccac tttccctgtg agcacactcc agatatacag 1921 agagccctca gcagagcctg ccgccacgta actgccatca gggctgaaga caactctggt 1981 ccagtcagag ccgcacttga acccaggtgc actgaatgtc tgcttgatag catttgttcg 2041 gagatcaata acttttagca agtcatcacg ggagcagctc aggagctcag tcctttctgg 2101 gtttaagtcc agggcagtaa tctttcccaa cagctccatc tctcgaacta tgctctctga 2161 gcgaatgtcc cagaaacgaa ttttcttgtc aaaatgtcca ctcattacac attgctctgt 2221 gcagacaata tcattgcaac tggatcctgc aaacactgtc tttatgcaga ctttgctgcg 2281 tagatcccag agtttgagag tccggtcgtg acttcctgag acaatccgcg cattgtccag 2341 caggaactta gcagacagca ctttcccact gtgtcccgtg agtgtgtgcc gtaatcgata 2401 atcatccaca gtccagattc ggcttgcaaa atcatttgaa gctgctaaga ggtaagatcc 2461 agcactatca aattcaatgc ttgtaattcc tgcattactg ccagataggg aacccttgaa 2521 ctcacatttt tctccaaata cttcccaaag cttaaccctg cggtccatgc ctccagtggc 2581 cagtaaccgg gaacctggac tgaactgcac agcgttgact tccccatcat gtgcatcgaa 2641 gacacacaag gcagtagctg gtaccctcac ttctttacca gaaccaggat gagtatccac 2701 attgtcctgg gggactggga aggaagagac agagcgtctc gtggctgctc tgctgatggc 2761 tcgcacagga gaggtctctt ctgtgtgatc agaagtttca tccacaatga cctcaatgtc 2821 atcatcctgt tcgactggta gaggttcctt tgctgcttct gcaagctctt tctgcagccg 2881 ggcttgccgc ctcctggagt ctttttcatt ctctgcatta agccgattgg cttcctgggc 2941 tttctcagcc atccatctgg tgaccagctc ctggttctct tccgtagttt tcctcagttt 3001 tccctccaag gcagtaaaag tgatctgcag ggcatcatat tcatccttca gggtctggtt 3061 ggctctttca aggtcacaaa gcttagtgcg caggtctagg cactccgtct ccaggtcaga 3121 gatagtctgc aaacattctg caatttttgc ttcattcatc tgcatctccc tgtccttccg 3181 ctgcatttgg ttattcaggt caatcaccag ttgagctaac tccccacgtt tcttgtgtaa 3241 ttcagtcagt tcctcttggt gcttaatcct cagttgggcc atttcttgta gctgattgtc 3301 attccatgtg ccatcatgtc cgggacttat ctcgtgcctg tttggtacgt catgcttttc 3361 agcctgtagt ttctgggcca acactgaatg aagatctgac ttttccagca atttgttata 3421 ctgcaggatg atctcctcga acgcctgtct ctgcagccgg tcccggcgcc tcagttgctc 3481 cgagatgtgg cgcttccagc gggggaagtc ggcggcgcgg aggcccgacg acatcgaatt 3541 caattcatcg atagatccgc cgccacccga cccaccaccg cccgagccac cgccaccctt 3601 gtacagctcg tccatgccga gagtcaagat cccggcggcg gtcacgaact ccagcaggac 3661 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg 3721 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc 3781 gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagtgggcct tgatgccgtt 3841 cttctgcttg tcggcggtga tatagacgtt gtggctgatg tagttgtact ccagcttgtg 3901 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag 3961 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa 4021 gatggtacgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt 4081 catgtggtcg gggtagcggc tgaagcactg cacgccccag gtcagggtgg tcacgagggt 4141 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta 4201 ggtggcatcg ccctcgccct cgccggacac gctgaacctg tggccgttta cgtcgccgtc 4261 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggc 4321 agatccttaa ttaactagct agatccggca gtctagagga tggtccaccc ccggggtcgg 4381 cagccttcac gtgggcggcg tgtatccaag ctgcgatgcc gtctactttg agggcggtgg 4441 gggtggtcag caggactgtg taaggtcctt tccagcgagg ttctaggttc ttagtctggt 4501 gtcggcggac ccacactgtg tcgccgactc ggtaagggtg aggtaccacc ggtcggtcca 4561 gttgttcttg gtaggctgcc gccagaggtc tccagacttc gtgctggact aagtagagag 4621 cctgtaagtg agcttggaga gaggggctgt tagtaactct tgtcatgtca gggtcaggga 4681 agtttacaag gggcgggggt gccccatata agatctcata tggccatatg ggggcgccta 4741 gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa 4801 gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct 4861 tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt 4921 ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc 4981 ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga 5041 aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat 5101 ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc gggcccatat 5161 tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg ctgcagcaga 5221 caagacgcgc ggcgcggctt cggtcccaaa ccgaaagcaa aaattcagac ggaggcggga 5281 actgttttag gttctcgtct cctaccagaa ccacatatcc ctcctctaag gggggtgcac 5341 caaagagtcc aaaacgatcg ggatttttgg actcaggtcg ggccacaaaa acggcccccg 5401 aagtccctgg gacgtctccc agggttgcgg ccgggtgttc cgaactcgtc agttccacca 5461 cgggtccgcc agatacagag ctagttagct aactagtacc gacgcaggcg cataaaatca 5521 gtcatagaca ctagacaatc ggacagacac agataagttg ctggccagct tacctcccgg 5581 tggtgggtcg gtggtccctg ggcaggggtc tcccgatccc ggacgagccc ccaaatgaaa 5641 gacccccgct gacgggtagt caatcactca gaggagaccc tcccaaggaa cagcgagacc 5701 acaagtcgga tgcaactgca agagggttta ttggatacac gggtacccgg gcgactcagt 5761 caatcggagg actggcgcgc cgagtgaggg gttgtgggct cttttattga gctcggggag 5821 cagaagcgcg cgaacagaag cgagaagcga actgattggt tagttcaaat aaggcacagg 5881 gtcatttcag gtccttgggg caccctggaa acatctgatg gttctctagc atggtaatag 5941 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg 6001 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat 6061 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc 6121 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc 6181 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact 6241 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata 6301 atcaatgtca acatggcggt aatgttggac atgagccaat ataaatgtac atattatgat 6361 atggatacaa cgtatgcaat gggccaagct cctcgagtga agacgaaagg gcctcgtgat 6421 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac 6481 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 6541 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 6601 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 6661 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 6721 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 6781 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 6841 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 6901 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 6961 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 7021 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 7081 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 7141 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 7201 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 7261 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 7321 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 7381 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 7441 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 7501 tttgcggccg gccgcaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 7561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 7621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 7681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 7741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 7801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 7861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 7921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 7981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 8041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 8101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 8161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 8221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 8281 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 8341 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 8401 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcaa ctatggccat 8461 ttaatgtaaa tacttaagaa aaaaaaccaa attaattttg atacatgctg catgtgaaag 8521 acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac agcgagacca 8581 caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg cgactcagtc 8641 aatcggagga ctggcgcccc gagtgagggg ttgtgggct //