LOCUS       sequence-376705-        5600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Test effect of extending a1/a2 on ADE2 editing rates in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5600)
  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 5600)
  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.040
FEATURES             Location/Qualifiers
     source          1..5600
                     /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(3133..3152)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3134..3152)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3162..3179)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3162..3178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3171..3193)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3319..3774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3406..3425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3616..3637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3905..4708)
                     /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(4709..4925)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4949..4968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5068..5090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5128..5146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5187..5600,1..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 tcgtcatacg gaatcttatc
     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 ggtgcgcatg ataagattcc
     2581 gtaattaacg aaattgcccc agtttcagag ctatgctgga aacagcatag caagttgaaa
     2641 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttgatggccg
     2701 gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca ccttcgggtg gcgaatggga
     2761 cttaaagaaa gtggaatatt cattcatatc atattttttc tattaactgc ctggtttctt
     2821 ttaaattttt tattggttgt cgacttgaac ggagtgacaa tatatatata tatatttaat
     2881 aatgacatca ttatctgtaa atctgattct taatgctatt ctagttatgt aagagtggtc
     2941 ctttccataa aaaaaaaaaa aaagaaaaaa gaattttagg aatacaatgc agcttgtaag
     3001 taaaatctgg aatattcata tcgccacaac ttcttatgct tataaaagca ctaatgcctg
     3061 aatttatgtt gaaaatatgt gtcacaaata aagaaactgt gacatctgac acatttccac
     3121 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     3181 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3241 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3301 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3361 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3421 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3481 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3541 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3601 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3661 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3721 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     3781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
     3841 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     3901 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     3961 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     4021 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     4081 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     4141 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     4201 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     4261 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     4321 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     4381 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     4441 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     4501 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     4561 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     4621 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     4681 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc
     4741 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     4801 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     4861 aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aaatgatgaa
     4921 ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4981 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5041 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5101 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5161 tcatgataat aatggtttct tagacggatc gcttgcctgt aacttacacg cgcctcgtat
     5221 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
     5281 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
     5341 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
     5401 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
     5461 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
     5521 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
     5581 ttcggaaaac aaaaactatt
//
