LOCUS Exported 6927 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Group II intron maturase from Eubacterium rectale (wild-type sequence). This is a highly processive reverse transcriptase that is called "MarathonRT" . ACCESSION . VERSION . KEYWORDS pET-6xHis-SUMO-MarathonRT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6927) AUTHORS Zhao C, Liu F, Pyle AM TITLE An ultraprocessive, accurate reverse transcriptase encoded by a metazoan group II intron. JOURNAL RNA. 2018 Feb;24(2):183-195. doi: 10.1261/rna.063479.117. Epub 2017 Nov 6. PUBMED 29109157 REFERENCE 2 (bases 1 to 6927) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6927 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 107..126 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 210..229 /label=T7 /note="T7 promoter, forward primer" promoter 210..228 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 229..253 /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)." RBS 268..290 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 310..327 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 337..354 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 361..654 /codon_start=1 /gene="S. cerevisiae SMT3 (truncated)" /product="cleavable ubiquitin-like protein tag" /label=SUMO /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG" CDS 2001..2018 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2071..2089) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2085..2132 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2169..2624 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2256..2275) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2466..2487 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(2716..3531) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 3439..3458 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3523..3542 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3653..4241 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4142..4161 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4395..4412 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4427..4566 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4492..4511) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4652..4674 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4668..4859) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5668..6750) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 6712..6731 /label=LacI-R /note="LacI, reverse primer" promoter complement(6751..6828) /gene="lacI" /label=lacI promoter ORIGIN 1 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga 61 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga 121 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt 181 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc 241 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg 301 ggcagcagcc atcatcatca tcatcacggc agcggcctgg tgccgcgcgg cagcgctagc 361 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 421 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 481 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 541 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 601 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtatggat 661 accagcaatc tgatggaaca aattctgagc agcgataatc tgaatcgtgc atatctgcag 721 gttgtgcgta ataaaggtgc cgaaggtgtt gatggtatga aatataccga actgaaagaa 781 cacctggcca aaaatggtga aaccattaaa ggtcagctgc gtacccgtaa atacaaaccg 841 cagcctgcac gtcgtgttga aattccgaaa ccggatggtg gtgttcgtaa tctgggtgtt 901 ccgaccgtta ccgatcgttt tattcagcag gcaattgcac aggttctgac cccgatttat 961 gaagaacagt ttcatgatca cagctatggc tttcgtccga atcgttgtgc acagcaggcc 1021 attctgaccg cactgaatat tatgaatgat ggcaatgatt ggatcgtgga tatcgacctg 1081 gaaaaattct ttgataccgt gaaccacgat aaactgatga ccctgattgg tcgtacaatc 1141 aaagatggtg atgtgattag catcgtgcgt aaatatctgg ttagcggcat tatgatcgat 1201 gatgaatatg aggatagcat tgttggtaca ccgcagggtg gtaatctgag tccgctgctg 1261 gccaatatta tgctgaatga actggacaaa gagatggaaa aacgcggtct gaattttgtt 1321 cgttatgccg atgattgcat tattatggtt ggtagcgaaa tgagcgcaaa tcgtgttatg 1381 cgtaatatta gccgcttcat cgaagaaaaa ctgggcctga aagttaacat gaccaaaagc 1441 aaagttgatc gtccgagtgg tctgaaatac ctgggttttg gtttctattt tgatccgcgt 1501 gcacaccagt ttaaagcaaa accgcatgca aaaagcgtgg ccaaattcaa aaaacgcatg 1561 aaagaactga cctgtcgtag ctggggtgtt agcaatagct ataaagtgga aaaactgaac 1621 cagctgattc gcggttggat caactatttc aaaattggca gcatgaaaac cctgtgcaaa 1681 gaactggata gccgtattcg ttatcgtctg cgtatgtgta tttggaaaca gtggaaaaca 1741 ccgcagaatc aagaaaaaaa tctggtgaaa ctgggcattg atcgtaatac cgcacgtcgc 1801 gttgcatata ccggtaaacg tattgcatat gtgtgcaata aaggcgcagt taatgttgcc 1861 attagcaata aacgtctggc aagctttggt ctgattagta tgctggatta ctacatcgaa 1921 aaatgcgtga cctgctaaag acaagcttag gtatttattc ggcgcaaagt gcgtcgggtg 1981 atgctgccaa cttagtcgag caccaccacc accaccactg agatccggct gctaacaaag 2041 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg 2101 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg 2161 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 2221 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 2281 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 2341 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 2401 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 2461 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 2521 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 2581 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg 2641 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 2701 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat 2761 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc 2821 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc 2881 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc 2941 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac 3001 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt 3061 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt 3121 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc 3181 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt 3241 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa 3301 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt 3361 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc 3421 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt 3481 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct 3541 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg 3601 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 3661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 3721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 3781 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact 3841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 3901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 3961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 4021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 4081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 4141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 4201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 4261 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 4321 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 4381 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt 4441 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg 4501 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg 4561 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 4621 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 4681 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc 4741 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt 4801 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact 4861 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag 4921 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag 4981 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc 5041 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg 5101 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca 5161 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg 5221 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag 5281 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg 5341 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga 5401 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag 5461 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc 5521 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag 5581 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg 5641 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 5701 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 5761 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg 5821 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg 5881 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac 5941 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag 6001 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg 6061 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg 6121 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga 6181 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac 6241 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga 6301 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg 6361 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac 6421 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc 6481 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag 6541 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg 6601 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga 6661 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc 6721 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg 6781 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac 6841 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga 6901 gcaccgccgc cgcaaggaat ggtgcat //