LOCUS       sequence-205790-        3003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 transcription template for Xi3eX.
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-Xi3eX
FEATURES             Location/Qualifiers
     source          1..3003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..35)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     135..154
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     348..370
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     362..379
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     363..379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     400..419
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        400..418
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(811..827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(811..827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(824..846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    835..851
                     /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(859..865,866..883,884..889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    904..925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1042..1059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1213..1801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1293..1312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1972..2787)
                     /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(1972..2787)
                     /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     2695..2714
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2788..2892)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2960..2978
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
       61 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      121 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
      181 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      241 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      301 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      361 ttgtaaaacg acggccagtg aattgacgcg tattgggatt aatacgactc actatagggc
      421 gaattcgagc tcactctctt ccgcatcgct gtctgcgagg taccctaccc cccgagtatg
      481 gatccctcta aaagcgggca tgacttctag agtagtccag ggtttccgag ggtttccgtc
      541 gacgatgtca gctcgtctcg agggcgtact aactgggccc cttcttcttt ttccctcagg
      601 tcctacacaa catactgcag gacaaactct tcgcggtctc tgcatgcgag ctcactctct
      661 tccgcatcgc tgtctgcgag gtaccctacc ccccgagtat ggatccctct aaaagcgggc
      721 atgacttcta gagtagtcca gggtttccga gggtttccga caattgcatg aagcttatcc
      781 caatggcgcg ccgagcttgg ctcgagcatg gtcatagctg tttcctgtgt gaaattgtta
      841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
      901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
      961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1021 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1081 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1621 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttagaaaaa
     1981 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     2041 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2101 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2161 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2221 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2281 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     2341 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     2401 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     2461 tacctggaat gctgttttcc cagggatcgc agtggtgagt aaccatgcat catcaggagt
     2521 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     2581 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     2641 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     2701 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     2761 agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta
     2821 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2881 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     2941 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     3001 tga
//
