LOCUS       sequence-376685-        6284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 RT and ncRNA in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6284)
  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 6284)
  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.027
FEATURES             Location/Qualifiers
     source          1..6284
                     /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(3818..3837)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3819..3837)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3847..3864)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3847..3863)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3856..3878)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4004..4459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4091..4110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4301..4322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4590..5393)
                     /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(5394..5609)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5633..5652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5752..5774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5812..5830)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5871..6284,1..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 cagacgatct
     2881 tacgctgtct gcccaatcaa tgaagaaggt cgtaaaggcg cgggatttct tgttttctat
     2941 catcccgtcc gagggctwgg 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 aaaaaaaaga aaaaagaatt
     3661 ttaggaatac aatgcagctt gtaagtaaaa tctggaatat tcatatcgcc acaacttctt
     3721 atgcttataa aagcactaat gcctgaattt atgttgaaaa tatgtgtcac aaataaagaa
     3781 actgtgacat ctgacacatt tccacgtacc caattcgccc tatagtgagt cgtattacgc
     3841 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3901 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3961 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     4021 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4081 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4141 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4201 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4261 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4321 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4381 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4441 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4501 cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     4561 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     4621 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     4681 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     4741 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     4801 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     4861 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     4921 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     4981 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     5041 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     5101 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     5161 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     5221 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     5281 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     5341 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     5401 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     5461 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     5521 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa
     5581 aaagaataaa aaaaaaatga tgaattgaaa aggtggtatg gtgcactctc agtacaatct
     5641 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5701 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5761 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5821 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg gatcgcttgc
     5881 ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt
     5941 gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga
     6001 gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac
     6061 tttacatata tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg
     6121 ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa
     6181 acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc
     6241 gatcccccta gagtctttta catcttcgga aaacaaaaac tatt
//
