LOCUS       sequence-376706-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 RT and ncRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  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 4682)
  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: pSLS.436
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1183..1200
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1301..1319)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1315..1362
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1592..2407)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1592..2407)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2315..2334
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2408..2499)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2515..3264)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2525..2542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2765..2784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3460..4542)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4504..4523
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4543..4620)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4666..4682,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4666..4682,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcatttaa ctatgataag attccgtatg cgcaccctta gcgagaggtt tatcattaag
       61 gtcaacctct ggatgttgtt tcggcatcct gcattgaatc tgagttactg tctgttttcc
      121 ttgttggaac ggagagcatc gcctgatgct ctccgagcca accaggaaac ccgttttttc
      181 tgacgtaagg gtgcgcaact ttcatgaaat ccgctgaata tttgaacact tttagattga
      241 gaaatctcgg cctacctgtc atgaacaatt tgcatgacat gtctaaggcg actcgcatat
      301 ctgttgaaac acttcggttg ttaatctata cagctgattt tcgctatagg atctacactg
      361 tagaaaagaa aggcccagag aagagaatga gaaccattta ccaaccttct cgagaactta
      421 aagccttaca aggatgggtt ctacgtaaca ttttagataa actgtcgtca tctccttttt
      481 ctattggatt tgaaaagcac caatctattt tgaataatgc taccccgcat attggggcaa
      541 actttatact gaatattgat ttggaggatt ttttcccaag tttaactgct aacaaagttt
      601 ttggagtgtt ccattctctt ggttataatc gactaatatc ttcagttttg acaaaaatat
      661 gttgttataa aaatctgcta ccacaaggtg ctccatcatc acctaaatta gctaatctaa
      721 tatgttctaa acttgattat cgtattcagg gttatgcagg tagtcggggc ttgatatata
      781 cgagatatgc cgatgatctc accttatctg cacagtctat gaaaaaggtt gttaaagcac
      841 gtgatttttt attttctata atcccaagtg aaggattggt tattaactca aaaaaaactt
      901 gtattagtgg gcctcgtagt cagaggaaag ttacaggttt agttatttca caagagaaag
      961 ttgggatagg tagagaaaaa tataaagaaa ttagagcaaa gatacatcat atattttgcg
     1021 gtaagtcttc tgagatagaa cacgttaggg gatggttgtc atttatttta agtgtggatt
     1081 caaaaagcca taggagatta ataacttata ttagcaaatt agaaaaaaaa tatggaaaga
     1141 accctttaaa taaagcgaag acctaatgac atgggcagca gccatcacca tcatcaccac
     1201 agccaggatc cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa
     1261 tgcttaagtc gaacagctta attaacctag gctgctgcca ccgctgagca ataactagca
     1321 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt
     1381 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
     1441 taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc
     1501 aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1561 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
     1621 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1681 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1741 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1801 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     1861 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1921 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc
     1981 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     2041 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     2101 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     2161 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     2221 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     2281 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     2341 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
     2401 ggctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgctcttccg
     2521 cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc
     2581 actcaaaagc ggtaatacgg ttatccacag aatcagggga taaagccgga aagaacatgt
     2641 gagcaaaaag caaagcaccg gaagaagcca acgccgcagg cgtttttcca taggctccgc
     2701 ccccctgacg agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga
     2761 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2821 ctgccgctta ccggatacat gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2881 agctcacgct gttggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2941 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3001 aacccggtaa gacacgactt atcgccactg gcagcagcca ttggtaactg atttagagga
     3061 ctttgtcttg aagttatgca cctgttaagg ctaaactgaa agaacagatt ttggtgagtg
     3121 cggtcctcca acccacttac cttggttcaa agagttggta gctcagcgaa ccttgagaaa
     3181 accaccgttg gtagcggtgg tttttcttta tttatgagat gatgaatcaa tcggtctatc
     3241 aagtcaacga acagctattc cgttactcta gatttcagtg caatttatct cttcaaatgt
     3301 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     3361 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     3421 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3541 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     3601 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     3661 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     3721 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     3781 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3841 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     3901 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     3961 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     4021 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     4081 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     4141 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     4201 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     4261 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     4321 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     4381 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     4441 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     4501 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     4561 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     4621 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
     4681 ta
//
