LOCUS       sequence-376707-        3448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 ncRNA library variants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3448)
  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 3448)
  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.601
FEATURES             Location/Qualifiers
     source          1..3448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     terminator      219..266
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(358..1173)
                     /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(358..1173)
                     /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     1081..1100
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1174..1265)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1281..2030)
                     /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(1291..1308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1531..1550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2226..3308)
                     /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
                     EACKAVVHNLLAQRVSGLIINYPLDDQDAIAVEAARTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHKQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3270..3289
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3309..3386)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3432..3448,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3432..3448,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccca tttaactatg ataagattcc gtatgcgcac
       61 ccttagcgag aggtttatca ttaaggtcaa cctctggatg ttgtttcggc atcctgcatt
      121 gaatctgagt tacaagctgt ttgtcgccag tcagactggc gacaacccgt tttttctgac
      181 gtaagggtgc gcaactttca tgaaatccgc tgcatcacct agcataaccc cttggggcct
      241 ctaaacgggt cttgaggggt tttttgaagt ggcacttttc ggggaaatgt gcgcggaacc
      301 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
      361 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
      421 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
      481 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
      541 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
      601 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
      661 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
      721 ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
      781 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
      841 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
      901 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
      961 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     1021 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     1081 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     1141 agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag
     1201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1261 acaaataggc atgcagcgct cttccgcttc ctcgctcact gactcgctac gctcggtcgt
     1321 tcgactgcgg cgagcggtgt cagctcactc aaaagcggta atacggttat ccacagaatc
     1381 aggggataaa gccggaaaga acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc
     1441 cgcaggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1501 gccagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacatgtcc gcctttctcc
     1621 cttcgggaag cgtggcgctt tctcatagct cacgctgttg gtatctcagt tcggtgtagg
     1681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1801 cagccattgg taactgattt agaggacttt gtcttgaagt tatgcacctg ttaaggctaa
     1861 actgaaagaa cagattttgg tgagtgcggt cctccaaccc acttaccttg gttcaaagag
     1921 ttggtagctc agcgaacctt gagaaaacca ccgttggtag cggtggtttt tctttattta
     1981 tgagatgatg aatcaatcgg tctatcaagt caacgaacag ctattccgtt actctagatt
     2041 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     2101 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     2161 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     2221 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2281 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     2341 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     2401 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     2461 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     2521 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     2581 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     2641 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     2701 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     2761 gcgatttgct tgtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     2821 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     2881 acattagtgc gggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     2941 atcagcccac tgacgcgttg cgcgagaaga ttgtgcacca ccgctttaca ggcttcgacg
     3001 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     3061 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     3121 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     3181 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     3241 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     3301 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     3361 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     3421 cattaggaaa ttaatacgac tcactata
//
