LOCUS       sequence-414426-        8221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains the R2Bm non-LTR retrotransposon 5'UTR and 3'UTR sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8221)
  AUTHORS   Wilkinson ME, Frangieh CJ, Macrae RK, Zhang F
  TITLE     Structure of the R2 non-LTR retrotransposon initiating target-primed
            reverse transcription.
  JOURNAL   Science. 2023 Apr 21;380(6642):301-308. doi: 
            10.1126/science.adg7883. Epub 2023 Apr 6.
  PUBMED    37023171
REFERENCE   2  (bases 1 to 8221)
  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"; doi: 
            "10.1126/science.adg7883"; journalName: "Science"; date: 
            "2023-04-21- 21"; volume: "380"; issue: "6642"; pages: "301-308"
COMMENT     Sequence Label: pMW37: pRS426-R2Bm RNA
FEATURES             Location/Qualifiers
     source          1..8221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      69..1411
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    401..448
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1438..1542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1543..1611,1612..2403)
                     /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     complement(1761..1780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2574..3162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3063..3082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3316..3333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3450..3471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3486..3491,3492..3509,3510..3516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3540
                     /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)."
     primer_bind     3529..3551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3548..3564
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3583..3603
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3585..3603
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    3666..3690
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    3821..3938
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4119..4142
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     misc_RNA        5807..5825
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     protein_bind    complement(6165..6189)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6237..6256)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6238..6256)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6266..6283)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6266..6282)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6275..6297)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6423..6878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6510..6529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6720..6741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7009..7812)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7813..8028)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8052..8071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8171..8193
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     3481 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3601 agggaacaaa agctggagct ccaccgcggt ggcggccccc gtcccaccat cgggcgcgga
     3661 tcatcacaag tttgtacaaa aaagcaggct ctttggccat ataggccacc aggtcctgca
     3721 ggcgcgctct tcaggaggcg cgccaactgc cctcttaatt aacgaagaag atcggaccgg
     3781 ggccgcggat ctttgcaaag atccaaaaaa agcatgcgta cggattagaa gccgccgagc
     3841 gggtgacagc cctccgaagg aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt
     3901 tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa caataaagat tctacaatac
     3961 tagcttttat ggttatgaag aggaaaattg gcagtaacct ggccccacaa acctcaaatg
     4021 aacgaatcaa attaacaacc agatatctcg actgaaaaaa aaggtttaaa ccagttccct
     4081 gaaattattc ccctacttga ctaataagta tataaagacg gtaggtattg attgtaattc
     4141 tgtaaatcta tttcttaaac ttcttgaatt ctacttttat agttagtctt ttttttagtt
     4201 ttaaaacacc aagaacttag tttcgaataa acacacataa ataaaccggc gggagtaact
     4261 atgactctct taaggggcga tacgcataat tttaattttt cgattcaaat ccagtcgtct
     4321 taatctggtg accagtggcg cggtcaccag tatagtgcac aggacgtgaa tggctccgag
     4381 gctggcggag tcactcacta taagtgtgag agacgatgtc ctgtgccaag tatacgtcca
     4441 accctaacgg gttaagtgaa attagttgct cataacaggg acggtgtacc tgtttgctcg
     4501 tggctggcta tcgaatggac gggaccaata cacccccctg ttagtaatgg ggtaagagag
     4561 agcggtctga aactatggcc gagatcacga cgccccactc ctacccataa cctgcacgtg
     4621 gtaccgccgc acattgaccg atacgggagg aggggcagca cttgaatcac gtagtcttgg
     4681 tgtagccatt gcgggactac agccctcgta agtgccgcct tagaacgcaa cggggcaata
     4741 ggtgggccgg ggcgctagcg ggggggagta atctcccctg ttggcgtgca ccgcactgct
     4801 ccctctgggg gcagtgtcat ccggaaacag gtgggccggg gcgccaccag gggggagcaa
     4861 tccctcctga tgatggcgag caccgcactg tcccttatgg gacggtgtaa cccggatggc
     4921 tgtacacgtg gtaaacacgt gacagcagcc ccgatggacg gaccgcgagg accgtcaagc
     4981 ctagcaggta ccttcgggtg gggccttgcg atacctgcgg gcgaaccctg tggtcgggtt
     5041 tgcagcccgg ccacagtggg tttttttcct gttgcaaaaa agtcaaataa agaaaataga
     5101 cctgaagcct ctggcctccc gctggagtca gagaggacag gcgataaccc gactgtgcgg
     5161 ggttccgccg gcgcagatcc tgtgggtcag gatgcgcctg gttggacctg ccagttctgc
     5221 gaacgaacct tttcgaccaa caggggtttg ggtgtccaca agcgtagagc ccaccctgtt
     5281 gagaccaata cggatgccgc tccgatgatg gtgaagcggc ggtggcatgg cgaggaaatc
     5341 gacctcctcg ctcgcaccga ggccaggttg ctcgctgagc ggggtcagtg ctcgggtgga
     5401 gacctctttg gcgcgcttcc agggtttgga agaactctgg aagcgattaa ggccttgcac
     5461 agtagtccag cggtaagggt gtagatcagg cccgtctgtt tctcccccgg agctcgctcc
     5521 cttggcttcc cttatatatt ttaacatcag aaacagacat taaacatcta ctgatccaat
     5581 ttcgccggcg tacggccacg atcgggaggg tgggaatctc gggggtcttc cgatcctaat
     5641 ccatgatgat tacgacctga gtcactaaag acgatggcat gatgatccgg cgatgaaaat
     5701 agccaaatgc ctcgtcatct aattagtgac gcgcatgaat ggcgtacacc atcagggtac
     5761 gagctagcca tatatgcacg agcatcagcc gtgcactagt agatctacat gaggatcacc
     5821 catgtcttaa gtcgactttg ttcccactgt acttttagct cgtacaaaat acaatatact
     5881 tttcatttct ccgtaaacaa catgttttcc catgtaatat ccttttctat ttttcgttcc
     5941 gttaccaact ttacacatac tttatatagc tattcacttc tatacactaa aaaactaaga
     6001 caattttaat tttgctgcct gccatatttc aatttgttat aaattcctat aatttatcct
     6061 attagtagct aaaaaaagat gaatgtgaat cgaatcctaa gagaattccc ctcgaagatc
     6121 gatctggcgg cccggccggc cgcggccgca ctcgagatat ctagacccag ctttcttgta
     6181 caaagtggtg atagcttgtc gagaagtact cgaggggggg cccggtaccc aattcgccct
     6241 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     6301 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     6361 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6421 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6661 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6781 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6841 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     6901 acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc
     6961 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     7021 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     7081 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     7141 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     7201 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     7261 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     7321 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     7381 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     7441 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     7501 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     7561 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     7621 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     7681 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     7741 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     7801 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
     7861 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     7921 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     7981 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg
     8041 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     8101 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8161 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     8221 a
//
