LOCUS       sequence-435227-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Ade2 and Faa1 editrons from a single Gal7 promoter, 
            separated by a Csy4 recognition sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Gonzalez-Delgado A, Lopez SC, Rojas-Montero M, Fishman CB, Shipman 
            SL
  TITLE     Simultaneous multi-site editing of individual genomes using retron 
            arrays.
  JOURNAL   Nat Chem Biol. 2024 Nov;20(11):1482-1492. doi: 
            10.1038/s41589-024-01665-7. Epub 2024 Jul 9.
  PUBMED    38982310
REFERENCE   2  (bases 1 to 5993)
  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/s41589-024-01665-7"; journalName: "Nat Chem Biol"; date: 
            "2024-11"; volume: "20"; issue: "11"; pages: "1482-1492"
COMMENT     Sequence Label: pSCL391
FEATURES             Location/Qualifiers
     source          1..5993
                     /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"
     protein_bind    2693..2712
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(3527..3546)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3528..3546)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3556..3573)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3556..3572)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3565..3587)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3713..4168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3800..3819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4010..4031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4299..5102)
                     /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(5103..5318)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5342..5361)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5461..5483
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5521..5539)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5580..5993,1..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 cctcaaaagg
     2281 gtgcgcatct gatgagtccg tgaggacgaa acgagctagc tcgtcatgat aagattccgt
     2341 atgcgcaccc ttagcgagag gtttatcatt aaggtcaacc tctggatgtt gtttcggcat
     2401 cctgcattga atctgagtta ctgtctgttt tccttggaaa tgttctattt agaaacaggg
     2461 gaattgctta ttaacgaaat tgcctgaagg cctcacaact ctggacatta taccattgat
     2521 gcttgcgtca cttcaggaaa cccgtttctt ctgacgtaag ggtgcgcata cggaatctta
     2581 tcaattaacg aaattgcccc agtttcagag ctatgctgga aacagcatag caagttgaaa
     2641 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttgttcactg
     2701 ccgtataggc agctaagaaa tgataagatt ccgtatgcgc acccttagcg agaggtttat
     2761 cattaaggtc aacctctgga tgttgtttcg gcatcctgca ttgaatctga gttactgtct
     2821 gttttcctca ggattcttta cctagcttca agatgtcgtc aaagctaaag gtcttgatat
     2881 ctcatctaac ttctttaatt ctgttgatgg catcatgagc agattgatag gaaacccgtt
     2941 tcttctgacg taagggtgcg catacggaat cttatcagct aaaggtcttg atatcgtttc
     3001 agagctatgc tggaaacagc atagcaagtt gaaataaggc tagtccgtta tcaacttgaa
     3061 aaagtggcac cgagtcggtg ctttttgatg gccggcatgg tcccagcctc ctcgctggcg
     3121 ccggctgggc aacaccttcg ggtggcgaat gggacttaaa gaaagtggaa tattcattca
     3181 tatcatattt tttctattaa ctgcctggtt tcttttaaat tttttattgg ttgtcgactt
     3241 gaacggagtg acaatatata tatatatatt taataatgac atcattatct gtaaatctga
     3301 ttcttaatgc tattctagtt atgtaagagt ggtcctttcc ataaaaaaaa aaaaaaagaa
     3361 aaaagaattt taggaataca atgcagcttg taagtaaaat ctggaatatt catatcgcca
     3421 caacttctta tgcttataaa agcactaatg cctgaattta tgttgaaaat atgtgtcaca
     3481 aataaagaaa ctgtgacatc tgacacattt ccacgtaccc aattcgccct atagtgagtc
     3541 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3601 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3661 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     3721 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3781 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3841 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3901 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3961 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4021 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4081 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4141 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     4201 gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt
     4261 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     4321 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     4381 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     4441 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     4501 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     4561 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     4621 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
     4681 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
     4741 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
     4801 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
     4861 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
     4921 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
     4981 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
     5041 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
     5101 atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg
     5161 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
     5221 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga
     5281 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca
     5341 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5401 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5461 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5521 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgg
     5581 atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat
     5641 ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa
     5701 ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa
     5761 aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata gatatacatt
     5821 cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga
     5881 caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta
     5941 tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact att
//
