LOCUS Exported 6204 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP-Atg14L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6204) AUTHORS Matsunaga K, Saitoh T, Tabata K, Omori H, Satoh T, Kurotori N, Maejima I, Shirahama-Noda K, Ichimura T, Isobe T, Akira S, Noda T, Yoshimori T TITLE Two Beclin 1-binding proteins, Atg14L and Rubicon, reciprocally regulate autophagy at different stages. JOURNAL Nat Cell Biol. 2009 Apr . 11(4):385-96. PUBMED 19270696 REFERENCE 2 (bases 1 to 6204) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6204 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 614..623 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 620..1336 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 620..1336 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(665..686) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(926..945) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1273..1294 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2999..3120 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3036..3055) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3090..3109 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3127..3582) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3264..3285) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3476..3495 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3609..3713 /gene="bla" /label=AmpR promoter promoter 3715..4072 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3738..3758) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3923..4058 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3985..4004 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4107..4901 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4161..4180) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4771..4790 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5089..5108) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5133..5180 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5509..6097 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5998..6017 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 1321 ggacgagctg tacaagtccg gccggactca gatctcgagc tcaagcttcg aattcatggc 1381 gtctcccagt gggaagggag cccgggcgct ggaggctcct ggctgcgggc cccggccgct 1441 cgcccgggac ctggtggact ccgtggacga tgcggagggg ctgtacgtgg ctgtggagcg 1501 ctgcccgctg tgcaacacta cccgccggcg gctgacctgc gccaaatgcg ttcagagcgg 1561 cgatttcgtc tacttcgacg gccgcgaccg ggagaggttt atcgacaaga aggaaaggct 1621 aagccgactt aagagcaagc aagaagaatt tcagaaagaa gtgttaaaag ctatggaagg 1681 aaaatggata acagatcagt tgagatggaa aataatgtcc tgcaagatga ggattgaaca 1741 gttaaaacaa acaatatgta aaggaaatga agaaatggag aaaaattctg aaggccttct 1801 caaaaccaag gaaaagaatc agaagcttta cagtcgagca caacggcacc aagagaaaaa 1861 ggagaagatt cagaggcata atcgcaaact tggtgacctg gtagaaaaaa agaccattga 1921 cttaagaagt cattatgagc gtctggcaaa tcttcgacga tcccatatat tagagctcac 1981 ctctgtcatt tttccaatcg aggaagtaaa gacgggtgtg agagaccccg cagatgtgtc 2041 ttcagagagt gacagtgcca tgacctccag cactgtgagc aagcttgctg aagcccggag 2101 gacaacttac ctctcaggac gatgggtctg tgacgatcac aacggagaca ccagcattag 2161 cattacaggg ccttggatta gcctccctaa caatggggac tactctgcct actacagctg 2221 ggtggaggag aagaaaacaa cccaggggcc tgacatggag cagagtaacc ctgcctacac 2281 catcagtgct gcgctgtgct atgcaactca gctggtcaac attctgtctc atatacttga 2341 tgtaaatctt cccaaaaagc tctgcaacag tgaattttgt ggcgaaaatc taagcaagca 2401 gaaatttact cgagcagtga agaaactgaa tgcaaatatt ctttaccttt gtttttctca 2461 gcatgtaaat ttagatcaat tacaaccact gcataccctc aggaatctaa tgtacctggt 2521 cagtccaagc tctgaacacc taggcaggtc agggcccttt gaagtacgag cagaccttga 2581 ggagtccatg gaatttgtgg atcccggagt tgctggagaa tcagatgaga gcggagatga 2641 gcgcgtcagc gatgaagaaa ccgacctggg cacagactgg gagaacttgc ctagtccccg 2701 gttttgtgat atcccttccc agtctgtgga agtctcccag agtcagagca cccaggcgtc 2761 cccacccatc gcgagcagca gtgcaggtgg gatgatctcc tctgcagcag cctcggtgac 2821 ctcctggttt aaagcttaca ctggacaccg ttaaggtacc gcgggcccgg gatccaccgg 2881 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 2941 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 3001 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 3061 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 3121 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 3181 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 3241 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 3301 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 3361 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 3421 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 3481 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 3541 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg 3601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 3661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg 3721 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 3901 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 3961 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga 4021 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc 4081 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 4141 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 4201 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 4261 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca 4321 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 4381 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 4441 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 4501 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 4561 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 4621 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 4681 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 4741 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 4801 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4861 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 4921 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 4981 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 5041 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa 5101 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca 5161 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga 5221 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca 5281 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc 5341 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa 5401 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 5461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 5521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 5581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 5641 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 5701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 5761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 5821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 5881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 5941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 6001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 6061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 6121 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 6181 ttctgtggat aaccgtatta ccgc //