LOCUS       sequence-376556-        5466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 RT in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5466)
  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 5466)
  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.006
FEATURES             Location/Qualifiers
     source          1..5466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             323..343
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1536..1555)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1537..1555)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1565..1582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1565..1581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1574..1596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1722..2177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1809..1828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2019..2040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2437..3099)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3100..3287)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(3434..3453)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3553..3575
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3613..3631)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3699..3803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3804..3872,3873..4664)
                     /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(4022..4041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4835..5423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5324..5343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggcacatct gcgtttcagg aacgcgaccg gtgaagacga ggacgcacgg aggagagtct
       61 tccgtcggag ggctgtcgcc cgctcggcgg cttctaatcc gtacttcaat atagcaatga
      121 gcagttaagc gtattactga aagttccaaa gagaaggttt ttttaggcta agataatggg
      181 gctctttaca tttccacaac atataagtaa gattagatat ggatatgtat atggtggtaa
      241 tgccatgtaa tatgattatt aaacttcttt gcgtccatcc aaaaaaaaag taagaatttt
      301 tgaaaattca atataaatgc caccaaagaa gaaaagaaag gtcaagtccg ctgaatactt
      361 gaacaccttc agattgagaa atttgggttt gccagtcatg aacaacttgc acgatatgtc
      421 taaggctacc agaatttctg ttgaaacctt gagattattg atctacaccg ccgatttcag
      481 atacagaatc tacactgttg aaaagaaggg tccagaaaag agaatgagaa ctatctacca
      541 accatccaga gaattgaagg ctttacaagg ttgggttttg agaaacatct tggacaagtt
      601 atcctcctcc ccattttcta tcggttttga aaagcaccaa tccatcttga acaatgccac
      661 tccacatatt ggtgccaact tcattttgaa catcgacttg gaagatttct tcccatcttt
      721 gactgctaac aaggttttcg gtgttttcca ttctttgggt tacaacagat tgatctcctc
      781 tgttttgacc aagatttgct gctacaagaa cttgttgcca caaggtgctc catcttctcc
      841 aaaattggct aacttgattt gctccaagtt ggactacaga attcaaggtt atgctggttc
      901 tagaggtttg atatacacta gatacgctga tgacttgacc ttgtctgctc aatctatgaa
      961 gaaagttgtt aaggccagag acttcttgtt ctccattatt ccatcagaag gtttggtcat
     1021 caactccaaa aagacctgta tttctggtcc aagatcccaa agaaaagtta ccggtttggt
     1081 tatctcccaa gaaaaggttg gtattggtag agaaaagtac aaagaaatca gagccaagat
     1141 ccaccatatt ttctgcggta aatcctccga aatcgaacat gttagaggtt ggttgtcttt
     1201 catcttgtcc gttgattcta agtcccacag aagattgatt acctacatct ctaagttgga
     1261 aaagaagtac ggtaagaacc cattgaacaa ggctaagact tgatttgcca gcttactatc
     1321 cttcttgaaa atatgcactc tatatctttt agttcttaat tgcaacacat agatttgctg
     1381 tataacgaat tttatgctat tttttaaatt tggagttcag tgataaaagt gtcacagcga
     1441 atttcctcac atgtagggac cgaattgttt acaagttctc tgtaccacca tggagacatc
     1501 aaaaattgaa aatctatgga aagatatgca attcgcccta tagtgagtcg tattacgcgc
     1561 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     1621 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1681 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     1741 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1801 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1861 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1921 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1981 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2041 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2101 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2161 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2221 caccgcatag atccgtcgag ttcaagagaa aaaaaaagaa aaagcaaaaa gaaaaaagga
     2281 aagcgcgcct cgttcagaat gacacgtata gaatgatgca ttaccttgtc atcttcagta
     2341 tcatactgtt cgtatacata cttactgaca ttcataggta tacatatata cacatgtata
     2401 tatatcgtat gctgcagctt taaataatcg gtgtcactac ataagaacac ctttggtgga
     2461 gggaacatcg ttggtaccat tgggcgaggt ggcttctctt atggcaaccg caagagcctt
     2521 gaacgcactc tcactacggt gatgatcatt cttgcctcgc agacaatcaa cgtggagggt
     2581 aattctgcta gcctctgcaa agctttcaag aaaatgcggg atcatctcgc aagagagatc
     2641 tcctactttc tccctttgca aaccaagttc gacaactgcg tacggcctgt tcgaaagatc
     2701 taccaccgct ctggaaagtg cctcatccaa aggcgcaaat cctgatccaa acctttttac
     2761 tccacgcacg gcccctaggg cctctttaaa agcttgaccg agagcaatcc cgcagtcttc
     2821 agtggtgtga tggtcgtcta tgtgtaagtc accaatgcac tcaacgatta gcgaccagcc
     2881 ggaatgcttg gccagagcat gtatcatatg gtccagaaac cctatacctg tgtggacgtt
     2941 aatcacttgc gattgtgtgg cctgttctgc tactgcttct gcctcttttt ctgggaagat
     3001 cgagtgctct atcgctaggg gaccaccctt taaagagatc gcaatctgaa tcttggtttc
     3061 atttgtaata cgctttacta gggctttctg ctctgtcatc tttgccttcg tttatcttgc
     3121 ctgctcattt tttagtatat tcttcgaaga aatcacatta ctttatataa tgtataattc
     3181 attatgtgat aatgccaatc gctaagaaaa aaaaagagtc atccgctagg tggaaaaaaa
     3241 aaaatgaaaa tcattaccga ggcataaaaa aatatagagt gtactagagg aggccaagag
     3301 taatagaaaa agaaaattgc gggaaaggac tgtgttatga cttccctgac taatgccgtg
     3361 ttcaaacgat acctggcagt gactcctagc gctcaccaag ctcttaaaac gggaatttat
     3421 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3481 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3541 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3601 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3661 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3781 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3841 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3901 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3961 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4021 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4081 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4141 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4201 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4261 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4321 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4381 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4441 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4501 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4561 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4621 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4681 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4741 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4921 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4981 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5041 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5101 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5161 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5221 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5281 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5341 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5401 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5461 tttgct
//
