LOCUS       pET-6xHis-SUMO-MarathonRT                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
            //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
//