LOCUS p1.1-Tr2-eGFP 7434 bp DNA circular SYN 28-OCT-2020 DEFINITION synthetic plasmid p1.1-Tr2-eGFP. ACCESSION p1.1-Tr2-eGFP VERSION KEYWORDS . SOURCE synthetic construct ORGANISM synthetic construct other sequences; artificial sequences. REFERENCE 1 (bases 1 to 7434) AUTHORS Orlova,N.A., Sinegubova,M.V. and Vorobiev,I.I. TITLE Optimization of the plasmid vector structures for stable and high-level expression in CHO cells JOURNAL unpublished REFERENCE 2 (bases 1 to 7434) AUTHORS Orlova,N.A., Sinegubova,M.V. and Vorobiev,I.I. TITLE Direct Submission JOURNAL Submitted (28-OCT-2020) lab of Mammalian Cells Bioengineering, Institute of Bioengineering of The Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, 60 let Oktjabrja pr-t, 7, bld. 1, Moscow, 117312, Russia COMMENT Bankit Comment: Vecscreen Comment:Submitter says that this sequence is a Synthetic Construct Bankit Comment: ALT EMAIL:ptichman@gmail.com Bankit Comment: TOTAL # OF SEQS:1. FEATURES Location/Qualifiers source 1..7434 /organism="synthetic construct" /mol_type="other DNA" /db_xref="taxon:32630" regulatory complement(150..823) /regulatory_class="replication_regulatory_region" /note="pUC replication origin" CDS complement(965..1825) /EC_number="3.5.2.6" /note="ampicillin resistance protein" /codon_start=1 /transl_table=11 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" regulatory complement(1820..1924) /regulatory_class="promoter" /note="bla promoter" regulatory complement(1960..2361) /regulatory_class="other" /note="EBVTR; fragment of the long terminal repeat from the Epstein-Barr virus" regulatory 2371..4882 /regulatory_class="other" /note="CHO EEF1A UFR; upstream flanking region of EEF1A1 gene" regulatory 3864..3869 /regulatory_class="TATA_box" CDS 4899..5618 /codon_start=1 /transl_table=11 /product="eGFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH MVLLEFVTAAGITLGMDELYK" regulatory 5640..6227 /regulatory_class="ribosome_binding_site" /note="EMCV IRES" CDS 6240..6803 /EC_number="1.5.1.3" /note="derived from Mus musculus" /codon_start=1 /transl_table=11 /product="dihydrofolate reductase" /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSV EGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIE QPELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLL PEYPGVLSEVQEEKGIKYKFEVYEKKD" regulatory 6805..7419 /regulatory_class="other" /note="CHO EEF1A DFR; downstream flanking region of EEF1A1 gene" regulatory 7070..7076 /regulatory_class="polyA_signal_sequence" BASE COUNT 1756 a 2007 c 2041 g 1630 t ORIGIN 1 cgtccggagc tcaggcgcgc catggctctt ccgcttcctc gctcactgac tcgctgcgct 61 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 721 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa 781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 1921 gggttccgcg cacatttccc cgaaaagtgc cacctgcatc tcatcacggt cacgcatggc 1981 tgcgcccgcg cagcgccccc cgaccccccc ccgatccccg gaacgccagc cgccatccac 2041 cgcccgcagc ccccggagcc cgcggacccc gaccccccgc cgctcgcccg ccgttgcgcc 2101 gctctctgcg ggggggctag ggggccgcgg ggggaaggcc acgccccctc cactttttcc 2161 aggaatgcgc ggcatgaccc gcccccccca tgacccgcca acaccccccc gatgcccccc 2221 gggggtcttt cctggggggc ctttgtcagg gttgcctgtg tcacccacgg gccccttggc 2281 cacgcttgcc tgggtcaccc cggagccccg ggtcacgttt gccggggtca gccctggggc 2341 gcttcttcac gctccattgc catgcagaat tcccacatgt ggacaccgga cagtaggtcc 2401 tcaaatgctc cttattaggt tggttcaata atatcaattg tttgttacta ggcagtgatg 2461 ttgtacatct ggaggagatc tcttgagccc ataatcaggt tattaggaat aaatactcta 2521 aggctaaaaa tgtagcttag tgataagagt gcttgcctgg tgtgctgaga ccctcggttc 2581 catctccaca accccatatt ccattacaaa ataccttttc accgtcccta gcattaagaa 2641 acaaaacaac aaagaagttt ttctttcttc tgagatcctg cccggagagg catttaaaac 2701 tggccagggc caaaaaaaaa aaaaaaaaaa gaaaaaaaag aaaagaaaac aggctagggc 2761 cggcatggtg gcgcacgcct ttaatcccag cacgcaggag gcagaggcag ggcggatctc 2821 tgtgagtttg aggtcagcct ggtctaccta gtgagtttca gggcacccag ggctaaagag 2881 actgtctcaa aaacaaaaca gccacacaat cagaaccaca gcaaaacgca gttatgatcc 2941 ttggaactgt aggaatgaca agcatttaaa taataggacg agccattttt gagaagctct 3001 gatttcacaa gtgtcaggga tgggctctgg gcgagtaaga ttgctaatgc tggcctctaa 3061 atgagaccac gtggagttga ttagattctt ttcatgttcc tcgtgctcta tcaaataact 3121 gtacccaaat acacacacac acacacacac acaatgcgcg cacacacaaa atcctttttt 3181 agcttaagaa gcccagaatc agaagtaaag ctaactgtgg gacttaagta ttattctgaa 3241 cggaactccc agggcgtgaa gcgcgcttca ggcttccaga gaagcagctg gcgctggatg 3301 gaatgaacca agaggccagc acaggggcag atccgtcgag ctctcggcca ccgagctgag 3361 cccttaggtt ctggggctgg gaagggtccc taggattgtg cacctctccc gcgggggaca 3421 agcaggggga tggcggggct gacgtcggga ggtggcctcc acgggaaggg acacccggat 3481 ctcgacacag ccttggcagt ggagtcagga agggtaggga cagattctgg acgccctctt 3541 ggccagctcc tcaccgcccc acccccgact ggagccgaga gtaattcata caaaaggagg 3601 gatcgccttc gcccctggga atcccaggga ccgtcgctaa attctggccg gcctcccagc 3661 ccggaaccgc tgtgcccgcc cagcgcggcg ggaggagcct gcgcctaggg cggatcgcgg 3721 gtcggcggga gagcacaagc ccacagtccc cggcggtggg ggaggggcgc gctgagcggg 3781 ggcccgggag ccagcgcggg gcaaactggg aaagtggtgt cgtgtgctgg ctccgccctc 3841 ttcccgaggg tgggggagaa cggtataaaa gtgcggtagt cgcgttggac gttctttttc 3901 gcaacgggtt tgccgtcaga acgcaggtga gtggcgggtg tggcctccgc gggcccgggc 3961 tccctccttt gagcggggtc ggaccgccgt gcgggtgtcg tcggccgggc ttctctgcga 4021 gcgttcccgc cctggatggc gggctgtgcg ggagggcgag ggggggaggc ctggcggcgg 4081 ccccggagcc tcgcctcgtg tcgggcgtga ggcctagcgt ggcttccgcc ccgccgcgtg 4141 ccaccgcggc cgcgctttgc tgtctgcccg gctgccctcg attgcctgcc cgcggcccgg 4201 gccaacaaag ggagggcgtg gagctggctg gtagggagcc ccgtagtccg catgtcgggc 4261 agggagagcg gcagcagtcg ggggggggac cgggcccgcc cgtcccgcag cacatgtccg 4321 acgccgcctg gacgggtagc ggcctgtgtc ctgataaggc ggccgggcgg tgggttttag 4381 atgccgggtt caggtggccc cgggtcccgg cccggtctgg ccagtacccc gtagtggctt 4441 agctccgagg agggcgaggc ccgcccgccc ggcaccagtt gcgtgcgcgg aaagatggcc 4501 gctcccgggc cctgtagcaa ggagctcaaa atggaggacg cggcagcccg gcggagcggg 4561 gcgggtgagt cacccacaca aaggaagagg gccttgcccc tcgccggccg ctgcttcctg 4621 tgaccccgtg gtgtaccggc cgcacttcag tcaccccggg cgctctttcg gagcaccgct 4681 ggcctccgct gggggagggg atctgtctaa tggcgttgga gtttgctcac atttggtggg 4741 tggagactgt agccaggcca gcctggccat gaaagtaatt cttggaattt gcccattttg 4801 agtttggagc gaagctgatt gacaaagctg cttagccgtt caaaggtatt cttcgaactt 4861 tttttttaag gtgttgtgaa aacctcgagg ccgccaccat ggtgagcaag ggcgaggagc 4921 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 4981 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 5041 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg 5101 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg 5161 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 5221 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 5281 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca 5341 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 5401 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 5461 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc 5521 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 5581 ccgggatcac tctcggcatg gacgagctgt acaagtaagc tagcggccgc gcagttaacg 5641 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg 5701 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc 5761 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa 5821 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag 5881 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg 5941 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg 6001 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa 6061 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg 6121 gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca 6181 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaaga tctgccacca 6241 tggttcgacc attgaactgc atcgtcgccg tgtcccaaaa tatggggatt ggcaagaacg 6301 gagacctacc ctggcctccg ctcaggaacg agttcaagta cttccaaaga atgaccacaa 6361 cctcttcagt ggaaggtaaa cagaatctgg tgattatggg taggaaaacc tggttctcca 6421 ttcctgagaa gaatcgacct ctaaaggaca gaattaatat agttctcagt agagaactca 6481 aagaaccacc acgaggagct cattttcttg ccaaaagttt ggatgatgcc ttaagactta 6541 ttgaacaacc ggaattggca agtaaagtag acatggtttg gatagtcgga ggcagttctg 6601 tttaccagga agccatgaat caaccaggcc acctcagact ctttgtgaca aggatcatgc 6661 aggaatttga aagtgacacg tttttcccag aaattgattt ggggaaatat aaacttctcc 6721 cagaataccc aggcgtcctc tctgaggtcc aggaggaaaa aggcatcaag tataagtttg 6781 aagtctacga gaagaaagac taatcatatt acccctaaca cctgccaccc cagtcttaat 6841 cagtggtgga agaacggtct cagaactgtt tgtctcaatt ggccatttaa gtttaatagt 6901 gaaagactgg ttaatgataa caatgcatcg gaaaaccttc aggaggaaag gagaatgttt 6961 tgtggaacat ttttgtgtgt gtggcagttt taagttatta gttttcaaaa tcagtacttt 7021 ttaatggaaa caacttgacc aaaaatctgt cacagaattt tgagacccat taaaatacaa 7081 gtttaatgag aagtctgtct ctgttaatgc tgaagtcatt actaagtgct tagcttagca 7141 aggtatgtgg atgcccattt gtgttccaag ggattggact gttcatcagg acccagagct 7201 gagtttcaag ggctcaagag atggcttatt acctgtgggt gtcttgaagg ttctggttgg 7261 gacaaattag gaatgttttt ggcagacatg gtgactaccc ttcatctggg tgagttcagt 7321 tgatttgtct tgagcctttg gggtttacac aagtaaatga catcatacag ttagtgtatt 7381 gttagtgaat attaatatat gaggcaggct ttgctctagc tagacatatg tcga //