LOCUS       sequence-368330-        8416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant B. mori truncated R2 RT expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8416)
  AUTHORS   Upton HE, Ferguson L, Temoche-Diaz MM, Liu XM, Pimentel SC, Ingolia 
            NT, Schekman R, Collins K
  TITLE     Low-bias ncRNA libraries using ordered two-template relay: Serial 
            template jumping by a modified retroelement reverse transcriptase.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Oct 19;118(42). pii: 2107900118. doi:
            10.1073/pnas.2107900118.
  PUBMED    34649994
REFERENCE   2  (bases 1 to 8416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2021-10-19"; volume: "118(42). pii: 
            2107900118. doi"; pages: " 10.1073/pnas.2107900118"
COMMENT     Sequence Label: 2Bc-T MBP_BoMoC(ed)_6xH
FEATURES             Location/Qualifiers
     source          1..8416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     421..438
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    453..592
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(518..537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     678..700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(694..885)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2343..2362
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2446..2465
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2446..2464
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2553..2575
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2584..3684
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALAAAQT"
     CDS             6223..6243
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6250..6267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6391..6409)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6405..6452
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6818..6823,6824..6840,6841..6846))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6873..6891)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6959..7063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7064..7132,7133..7924)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7282..7301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8095..8416,1..267)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
       61 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      301 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      361 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      421 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
      481 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
      541 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
      601 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
      661 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
      721 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
      781 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
      841 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
      901 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
      961 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     1021 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     1081 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     1141 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     1201 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     1261 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     1321 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
     1381 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
     1441 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     1501 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     1561 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     1621 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     1681 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     1741 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     1801 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     1861 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     1921 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     1981 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     2041 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     2101 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta
     2161 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     2221 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac gcggcctgcc
     2281 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     2341 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     2401 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     2461 tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt aattaagcgg
     2521 ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa ggagatatag
     2581 ttcatgaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
     2641 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
     2701 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
     2761 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
     2821 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
     2881 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
     2941 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
     3001 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
     3061 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
     3121 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
     3181 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
     3241 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
     3301 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
     3361 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
     3421 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
     3481 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc
     3541 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
     3601 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
     3661 gaagccctgg cagccgcgca gactgctgcg gccgcagcta tgggtcgtcg tcgtgcgtgc
     3721 cgtgcgatgc gtccgaagac cgcgggtcgt cgtaacgacc tgcacgacga tcgtaccgcg
     3781 agcgcgcaca agaccagccg tcagaaacgt cgtgcggagt acgcgcgtgt gcaagaactg
     3841 tataagaaat gccgtagccg tgcggcggcg gaagtgatcg atggtgcgtg cggtggcgtt
     3901 ggtcacagcc tggaggaaat ggaaacctac tggcgtccga ttctggaacg tgtgagcgac
     3961 gcgccgggtc cgaccccgga ggcgctgcac gcgctgggtc gtgcggaatg gcacggtggc
     4021 aaccgtgatt atacccagct gtggaagccg atcagcgttg aggaaattaa agcgagccgt
     4081 ttcgactggc gtaccagccc gggtccggat ggtatccgta gcggccagtg gcgtgcggtg
     4141 ccggttcacc tgaaggcgga aatgttcaac gcgtggatgg cgcgtggcga gatcccggaa
     4201 attctgcgtc aatgccgtac cgtgtttgtt ccgaaagttg agcgtccggg tggcccgggt
     4261 gaataccgtc cgatcagcat tgcgagcatc ccgctgcgtc acttccacag cattctggcg
     4321 cgtcgtctgc tggcgtgctg cccgccggac gcgcgtcagc gtggctttat ctgcgcggat
     4381 ggtaccctgg agaacagcgc ggtgctggac gcggttctgg gtgatagccg taagaaactg
     4441 cgtgaatgcc acgtggcggt tctggacttc gcgaaggcgt ttgataccgt gagccacgag
     4501 gcgctggttg aactgctgcg tctgcgtggc atgccggagc agttctgcgg ttacattgcg
     4561 cacctgtatg acaccgcgag caccaccctg gcggtgaaca acgaaatgag cagcccggtg
     4621 aaagttggcc gtggtgttcg tcaaggcgac ccgctgagcc cgatcctgtt taacgtggtt
     4681 atggatctga ttctggcgag cctgccggag cgtgtgggtt accgtctgga gatggaactg
     4741 gttagcgcgc tggcgtatgc ggacgatctg gtgctgctgg cgggcagcaa ggttggtatg
     4801 caggaaagca tcagcgcggt ggactgcgtt ggccgtcaaa tgggtctgcg tctgaactgc
     4861 cgtaaaagcg cggtgctgag catgatcccg gatggtcacc gtaagaaaca ccactacctg
     4921 accgagcgta ccttcaacat tggtggcaag ccgctgcgtc aggtgagctg cgttgaacgt
     4981 tggcgttatc tgggcgtgga ctttgaggcg agcggttgcg ttaccctgga acacagcatc
     5041 agcagcgcgc tgaacaacat tagccgtgcg ccgctgaaac cgcagcaacg tctggagatc
     5101 ctgcgtgcgc acctgattcc gcgtttccag cacggctttg ttctgggtaa catcagcgac
     5161 gatcgtctgc gtatgctgga tgtgcagatt cgtaaggcgg ttggtcaatg gctgcgtctg
     5221 ccggcggacg tgccgaaagc gtactatcac gcggcggttc aagatggtgg cctggcgatc
     5281 ccgagcgtgc gtgcgaccat cccggacctg attgttcgtc gttttggtgg cctggatagc
     5341 agcccgtgga gcgtggcgcg tgcggcggcg aagagcgaca aaattcgtaa gaaactgcgt
     5401 tgggcgtgga agcagctgcg tcgtttcagc cgtgtggata gcaccaccca acgtccgagc
     5461 gttcgtctgt tttggcgtga gcacctgcac gcgagcgttg acggtcgtga gctgcgtgaa
     5521 agcacccgta ccccgaccag caccaaatgg attcgtgaac gttgcgcgca gattaccggt
     5581 cgtgatttcg tgcaatttgt tcacacccac atcaacgcgc tgccgagccg tattcgtggc
     5641 agccgtggcc gtcgtggtgg cggtgagagc agcctgacct gccgtgcggg ttgcaaagtg
     5701 cgtgaaacca ccgcgcacat cctgcagcaa tgccaccgta cccacggcgg tcgtatcctg
     5761 cgtcacaaca agattgtgag cttcgttgcg aaggcgatgg aggaaaacaa atggaccgtg
     5821 gagctggaac cgcgtctgcg taccagcgtt ggcctgcgta aaccggcgat cattgcgagc
     5881 cgtgatggcg tgggtgttat cgtggacgtt caggtggtta gcggtcaacg tagcctggat
     5941 gagctgcacc gtgaaaagcg taacaaatac ggcaaccacg gtgagctggt tgagctggtt
     6001 gcgggccgtc tgggtctgcc gaaagcggag tgcgtgcgtg cgaccagctg caccattagc
     6061 tggcgtggcg tttggagcct gaccagctat aaagagctgc gtagcatcat tggtctgcgt
     6121 gaaccgaccc tgcagatcgt gccgattctg gcgctgcgtg gcagccacat gaactggacc
     6181 cgttttaacc aaatgaccag cgtgatgggt ggcggtgttg gtgagaacct ctacttccaa
     6241 tccggctctc atcaccatca ccatcactaa taaccaactc cataaggatc cgcgatcgcg
     6301 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
     6361 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     6421 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggacatc
     6481 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
     6541 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
     6601 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
     6661 tggacgacat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
     6721 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     6781 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
     6841 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6901 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6961 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7021 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
     7081 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7141 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7201 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7261 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     7321 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7381 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7441 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7501 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7561 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     7621 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7681 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7741 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7801 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7861 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7921 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7981 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8041 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8101 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8161 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8221 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     8281 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8341 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8401 gcggtcgggc tgaacg
//
