LOCUS       sequence-205789-        3003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 transcription template for Xi3e5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3003)
  AUTHORS   Braun JE, Friedman LJ, Gelles J, Moore MJ
  TITLE     Synergistic assembly of human pre-spliceosomes across introns and 
            exons.
  JOURNAL   Elife. 2018 Jun 22;7. pii: 37751. doi: 10.7554/eLife.37751.
  PUBMED    29932423
REFERENCE   2  (bases 1 to 3003)
  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"; journalName: "Elife."; 
            date: "2018-06-22"; volume: "7. pii: 37751. doi"; pages: " 
            10.7554/eLife.37751"
COMMENT     Sequence Label: T7-Xi3e5
FEATURES             Location/Qualifiers
     source          1..3003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..27
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(65..87)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     187..206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     400..422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     414..431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     415..431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     452..471
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        452..470
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(863..879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(863..879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(876..898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    887..903
                     /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)."
     promoter        complement(join(911..917,918..935,936..941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    956..977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1094..1111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1265..1853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1345..1364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2024..2839)
                     /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(2024..2839)
                     /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     2747..2766
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2840..2944)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
       61 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      121 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
      181 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
      241 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
      301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      361 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      421 cgacggccag tgaattgacg cgtattggga ttaatacgac tcactatagg gcgaattcga
      481 gctcactctc ttccgcatcg ctgtctgcga ggtaccctac cccccgagta tggatccctc
      541 taaaagcggg catgacttct agagtagtcc agggtttccg agggtttccg tcgacgatgt
      601 cagctcgtct cgagggcgta ctaactgggc cccttcttct ttttccctca ggtcctacac
      661 aacatactgc aggacaaact cttcgcggtc tctgcatgcg agctcactct cttccgcatc
      721 gctgtctgcg aggtacccta ccaggtgagt atggatccct ctaaaagcgg gcatgacttc
      781 tagagtagtc cagggtttcc gagggtttcc gacaattgca tgaagcttat cccaatggcg
      841 cgccgagctt ggctcgagca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
      901 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      961 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1021 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1081 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1141 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1201 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1261 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1321 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1381 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1441 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1501 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1561 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1621 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1681 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1741 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1801 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1861 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1921 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1981 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttagaaa aactcatcga
     2041 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2101 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2161 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2221 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     2281 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     2341 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     2401 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2461 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2521 atgctgtttt cccagggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2581 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2641 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2701 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2761 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     2821 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2941 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3001 taa
//
