LOCUS       sequence-205788-        3003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 transcription template for 5i3eX.
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: T7-5i3eX
FEATURES             Location/Qualifiers
     source          1..3003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     250..272
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     264..281
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     265..281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     302..321
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        302..320
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(713..729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(713..729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(726..748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    737..753
                     /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(761..767,768..785,786..791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    806..827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(944..961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1115..1703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1195..1214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1874..2689)
                     /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(1874..2689)
                     /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     2597..2616
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2690..2794)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2862..2880
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2918..2940)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
       61 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      121 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      181 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      241 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgacg cgtattggga
      301 ttaatacgac tcactatagg gcgaattcga gctcactctc ttccgcatcg ctgtctgcga
      361 ggtaccctac caggtgagta tggatccctc taaaagcggg catgacttct agagtagtcc
      421 agggtttccg agggtttccg tcgacgatgt cagctcgtct cgagggcgta ctaactgggc
      481 cccttcttct ttttccctca ggtcctacac aacatactgc aggacaaact cttcgcggtc
      541 tctgcatgcg agctcactct cttccgcatc gctgtctgcg aggtacccta ccccccgagt
      601 atggatccct ctaaaagcgg gcatgacttc tagagtagtc cagggtttcc gagggtttcc
      661 gacaattgca tgaagcttat cccaatggcg cgccgagctt ggctcgagca tggtcatagc
      721 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      781 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      841 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      901 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      961 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1021 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1081 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1141 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1201 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1261 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1321 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1381 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1441 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1501 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1561 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1621 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1681 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1861 cttggtctga cagttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1921 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1981 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     2041 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     2101 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     2161 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     2221 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     2281 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     2341 ctgaatcagg atattcttct aatacctgga atgctgtttt cccagggatc gcagtggtga
     2401 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     2461 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     2521 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     2581 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     2641 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt
     2701 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2761 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2821 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2881 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2941 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3001 cag
//
