LOCUS       sequence-376554-        6283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 dead RT and ncRNA in yeasst.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  AUTHORS   Lopez SC, Crawford KD, Lear SK, Bhattarai-Kline S, Shipman SL
  TITLE     Precise genome editing across kingdoms of life using retron-derived 
            DNA.
  JOURNAL   Nat Chem Biol. 2022 Feb;18(2):199-206. doi: 
            10.1038/s41589-021-00927-y. Epub 2021 Dec 23.
  PUBMED    34949838
REFERENCE   2  (bases 1 to 6283)
  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: 
            "10.1038/s41589-021-00927-y"; journalName: "Nat Chem Biol"; date: 
            "2022-02"; volume: "18"; issue: "2"; pages: "199-206"
COMMENT     Sequence Label: pSCL.037
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..231
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(232..300,301..1092)
                     /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(450..469)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1263..1851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1752..1771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3817..3836)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3818..3836)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3846..3863)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3846..3862)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3855..3877)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4003..4458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4090..4109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4300..4321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4589..5392)
                     /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(5393..5608)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5632..5651)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5751..5773
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5811..5829)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5870..6283,1..90)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttttctttaa tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt
       61 taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga
      121 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      181 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      241 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      301 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      361 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      421 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      481 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      541 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      601 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      661 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      721 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      781 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      841 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      901 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      961 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1021 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1081 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1141 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1261 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1321 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1381 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1441 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1501 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1561 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1621 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1681 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1741 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1801 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaaggccagc
     1861 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctggcct tttgctcaag
     1921 tagagggggt aatttttccc ctttattttg ttcatacatt cttaaattgc tttgcctctc
     1981 cttttggaaa gctatacttc ggagcactgt tgagcgaagg ctcattagat atattttctg
     2041 tcattttcct taacccaaaa ataagggaaa gggtccaaaa agcgctcgga caactgttga
     2101 ccgtgatccg aaggactggc tatacagtgt tcacaaaata gccaagctga aaataatgtg
     2161 tagctatgtt cagttagttt ggctagcaaa gatataaaag caggtcggaa atatttatgg
     2221 gcattattat gcagagcatc aacatgataa aaaaaaacag ttgaatattc cctcaaaacg
     2281 ccacatgaaa tctgcagagt atctgaatac gttccgcctt aggaatttgg gcctccccgt
     2341 gatgaacaat ctccacgata tgagcaaggc gactcgaata tccgtggaaa cgctgagact
     2401 gctcatctat acagcagact ttcggtacag gatctacacg gtcgaaaaga aggggcctga
     2461 gaaacgcatg cgaacaattt atcaacctag ccgagagctc aaggcgttgc agggctgggt
     2521 tcttcgaaac atccttgaca aactctcatc atcacccttt agtattgggt ttgaaaagca
     2581 ccaaagcatc cttaacaacg cgacgccaca cataggtgcc aatttcatat tgaacatcga
     2641 cttggaggat ttttttccga gcctcacagc caataaagtg ttcggtgttt ttcacagtct
     2701 tgggtacaat cgccttatta gttccgttct taccaagatt tgttgttaca agaatctctt
     2761 gccccaggga gcacccagca gtccgaaatt ggcgaatttg atttgttcca agctcgatta
     2821 tcgaatacaa gggtacgcgg gcagccgggg actcatctat acccgctacg cagctgctct
     2881 tacgctgtct gcccaatcaa tgaagaaggt cgtaaaggcg cgggatttct tgttttctat
     2941 catcccgtcc gagggcttgg taattaattc caaaaagact tgtatctcag gaccacgatc
     3001 tcagcgaaaa gtgacaggac tcgtcatttc tcaagaaaaa gtcggtatag ggagagagaa
     3061 gtataaggaa atccgcgcga agatccacca catattctgt ggcaagagca gcgagataga
     3121 acacgtccga ggctggttgt ccttcatact gagcgtggac tcaaaaagcc accgccggtt
     3181 gatcacctat atttcaaaac tggaaaagaa atatggaaag aacccactca acaaagctaa
     3241 aacatagacc ggttagatgc gcacccttag cgagaggttt atcattaagg tcaacctctg
     3301 gatgttgttt cggcatcctg cattgaatct gagttactgt ctgttttcct tgttggaacg
     3361 gagagcatcg cctgatgctc tccgagccaa ccaggaaacc cgttatttct gacgtaaggg
     3421 tgcgcaaccg gttagtaatg agtttaaaca aagaaagtgg aatattcatt catatcatat
     3481 tttttctatt aactgcctgg tttcttttaa attttttatt ggttgtcgac ttgaacggag
     3541 tgacaatata tatatatata tttaataatg acatcattat ctgtaaatct gattcttaat
     3601 gctattctag ttatgtaaga gtggtccttt ccataaaaaa aaaaaaagaa aaaagaattt
     3661 taggaataca atgcagcttg taagtaaaat ctggaatatt catatcgcca caacttctta
     3721 tgcttataaa agcactaatg cctgaattta tgttgaaaat atgtgtcaca aataaagaaa
     3781 ctgtgacatc tgacacattt ccacgtaccc aattcgccct atagtgagtc gtattacgcg
     3841 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3901 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3961 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     4021 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4081 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4141 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4201 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4261 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4321 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4381 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4441 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4501 acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     4561 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     4621 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     4681 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     4741 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     4801 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     4861 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     4921 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     4981 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     5041 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     5101 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     5161 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     5221 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     5281 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     5341 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     5401 ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     5461 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     5521 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     5581 aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca gtacaatctg
     5641 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5701 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5761 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5821 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgg atcgcttgcc
     5881 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
     5941 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
     6001 ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact
     6061 ttacatatat atttattaga caagaaaagc agattaaata gatatacatt cgattaacga
     6121 taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa
     6181 caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg
     6241 atccccctag agtcttttac atcttcggaa aacaaaaact att
//
