LOCUS Exported 9527 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMXs-IP GFP-NBR1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9527) AUTHORS Itakura E, Mizushima N TITLE p62 Targeting to the autophagosome formation site requires self-oligomerization but not LC3 binding. JOURNAL J Cell Biol. 2011 Jan 10;192(1):17-27. PUBMED 21220506 REFERENCE 2 (bases 1 to 9527) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9527 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..375 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 313..329 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS 407..1123 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 407..1123 /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(452..473) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(713..732) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1060..1081 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4153..4739 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4320..4337) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4547..4566 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4740..5342 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 5239..5259 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5513..5982 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(6112..6129) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6283..6871) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6363..6382) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7035..7895) /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 7658..7677 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7896..8000) /gene="bla" /label=AmpR promoter LTR 8030..8623 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 8686..9043 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 9102..9518 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 9448..9470 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 9486..9502 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" ORIGIN 1 ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga ccctgacatg 61 acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta cttagtccag 121 cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg accggtggta 181 cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac taagaaccta 241 gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc cctcaaagta 301 gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga 361 ccatcctcta gactgccgga tctagctagt taattaagga tctgccatgg tgagcaaggg 421 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 481 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 541 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 601 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt 661 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 721 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 781 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 841 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa 901 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca 961 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca 1021 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 1081 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtggcg gtggctcggg 1141 cggtggtggg tcgggtggcg gcggatctat cgatgaattg aattcacgcg ttcgcgagtt 1201 aacggatctt atggaaccac aggttactct aaatgtgact tttaaaaatg aaattcaaag 1261 ctttctggtt tctgatccag aaaatacaac ttgggctgat atcgaagcta tggtaaaagt 1321 ttcatttgat ctgaatacta ttcaaataaa atacctggat gaggaaaatg aagaggtatc 1381 catcaacagt caaggagaat atgaagaagc gcttaagatg gcagttaaac agggaaacca 1441 actgcagatg caagtccacg aagggcacca tgtcgttgat gaagccccac ccccagttgt 1501 aggagcaaaa cgactagctg ccagggcagg gaagaagcca cttgcacatt actcttcact 1561 ggtgagagtc ttgggatcag acatgaagac cccagaggat cctgcagtgc agtcgtttcc 1621 acttgttcca tgtgacacag accagcctca agacaagccc ccagactggt tcacaagcta 1681 cctggagacg ttcagagaac aagtggttaa cgaaacggtt gagaagcttg aacagaaatt 1741 acatgaaaag cttgtcctcc agaacccatc cttgggttct tgtccctcag aagtctcaat 1801 gcctacttca gaagaaacat tgtttttgcc agaaaaccag ttcagctggc atattgcttg 1861 caacaactgc caaagaagga ttgttggtgt ccgctaccag tgtagcctat gcccatccta 1921 caatatctgt gaagattgtg aagcagggcc atatggccat gacactaacc acgtcctgct 1981 gaagttgcgg agacctgttg tgggctcctc tgaaccgttc tgtcactcaa agtactctac 2041 tcctcgtctt cctgctgctc tggaacaagt caggctccag aaacaggttg ataagaactt 2101 tcttaaagca gaaaagcaaa ggttgcgagc tgagaagaaa caacgtaaag cagaggtcaa 2161 ggaacttaaa aagcagctta aactccatag gaaaattcac ctgtggaatt caatccatgg 2221 actccagagc cccaagtctc ctttaggccg acctgagagc ttgctccagt ctaataccct 2281 gatgctccct ttgcagccct gtacctccgt tatgccaatg ctcagtgcag catttgtgga 2341 tgagaatttg cctgatggga ctcaccttca gccaggaacc aagtttatca aacactggag 2401 gatgaaaaat acaggaaatg taaagtggag tgcagacaca aagctcaagt tcatgtgggg 2461 aaacctgact ttggcttcca cagaaaagaa ggatgttttg gttccctgcc tcaaggccgg 2521 ccatgtggga gttgtatctg tggagttcat tgccccagcc ttggagggaa cgtatacttc 2581 ccattggcgt ctttctcaca aaggccagca atttgggcct cgggtctggt gcagtatcat 2641 agtagatcct ttcccctccg aagagagccc tgataacatt gaaaagggca tgatcagctc 2701 aagcaaaact gatgatctca cctgccagca agaggaaact tttcttctgg ctaaagaaga 2761 aagacagctt ggtgaagtga ctgagcagac agaagggaca gcagcctgca tcccacagaa 2821 ggcaaaaaat gttgccagtg agagggagct ctacatccca tctgtggatc ttctgactgc 2881 ccaggacctg ctgtcctttg agctgttgga tataaacatt gttcaagagt tggagagagt 2941 gccccacaac acccctgtgg atgtgactcc ctgcatgtct cctctgccac atgacagtcc 3001 tttaatagag aagccaggct tggggcagat agaggaagag aatgaagggg caggatttaa 3061 agcacttcct gattctatgg tgtcagtaaa gaggaaggct gagaacattg cttctgtgga 3121 ggaagcagaa gaagacctga gtgggaccca gtttgtgtgt gagacagtaa tccgatccct 3181 taccttggat gctgccccag accacaaccc tccttgcaga cagaagtcct tgcagatgac 3241 atttgccttg cctgaaggac cacttggaaa tgagaaggag gagattatcc atatcgctga 3301 ggaagaagct gtcatggagg aggaggagga tgaggaggat gaggaggagg aggatgagct 3361 caaagatgaa gttcaaagtc agtcctctgc ttcctcagag gattacatca tcatcctgcc 3421 tgagtgcttt gataccagcc gccccctggg ggattctatg tacagctctg cgctctcaca 3481 gccaggcctg gagcgaggtg ctgaaggcaa gcctggggtt gaggctgggc aggaaccagc 3541 tgaggctggg gaaagactcc ctggagggga gaaccagcca caggagcaca gcataagtga 3601 catcctcacg acctcacaga ctctggaaac agtgccccta atcccagagg tagtggagct 3661 tccaccgtca ctgcccagga gctctccttg tgtacatcat catggttccc caggagtgga 3721 tttaccagtt accataccag aagtttcttc agtccctgat cagatcagag gagagcccag 3781 aggctcatca ggacttgtaa acagcagaca gaagagctat gaccactcaa ggcaccatca 3841 tgggagcagc attgctggag gactggtgaa gggggctttg tctgttgctg cctctgcata 3901 caaggccctg tttgctgggc caccagtcac tgcacagcca ataatttctg aagatcagac 3961 agcagccctg atggcccatc tctttgaaat gggattctgt gacaggcagc tgaacctacg 4021 gctgctgaag aaacacaatt acaatatcct gcaggttgtg acagaacttc ttcagttaaa 4081 caacaacgac tggtacagcc aacgctattg aagatccctc gagcggccgc tacgtaaatt 4141 ccgccccccc cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga 4201 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa 4261 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc 4321 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc 4381 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg 4441 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca 4501 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag 4561 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct 4621 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc 4681 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca 4741 tggttaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg 4801 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 4861 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 4921 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 4981 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 5041 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 5101 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 5161 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 5221 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 5281 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 5341 gagtcgacga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac 5401 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat 5461 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat 5521 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat 5581 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 5641 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat 5701 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc 5761 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc 5821 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg 5881 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg 5941 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa 6001 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat 6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6961 gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa 7021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 7081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 7141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 7201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 7261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 7321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 7381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 7441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 7501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 7561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 7621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 7681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 7801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcata atgaaagacc 8041 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata 8101 cataactgag aatagaaaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg 8161 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg 8221 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag 8281 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc 8341 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca 8401 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc 8461 cacaacccct cactcggcgc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt 8521 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt 8581 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga 8641 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac 8701 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta 8761 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttcggaac 8821 acccggccgc aaccctggga gacgtcccag ggacttcggg ggccgttttt gtggcccgac 8881 ctgagtccaa aaatcccgat cgttttggac tctttggtgc acccccctaa taggagggat 8941 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc 9001 tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt 9061 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc 9121 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg 9181 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg 9241 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg 9301 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag 9361 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc 9421 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat 9481 cctcccttta tccagccctc actccttctc taggcgcccc catatgg //