LOCUS       sequence-240714-        3376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 transcription template for 5iX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3376)
  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 3376)
  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: T7-5iX
FEATURES             Location/Qualifiers
     source          1..3376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(261..279)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(300..316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(300..316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(313..335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    324..340
                     /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(348..354,355..372,373..378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    393..414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(531..548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(702..1290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(782..801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1461..2252,2253..2321))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2084..2103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2322..2426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2494..2512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2550..2572)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2672..2691
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      2758..3213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2845..2864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3055..3076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3339..3361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3353..3370
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3354..3370
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattcgag ctcactctct tccgcatcgc tgtctgcgag
       61 gtaccctacc aggtgagtat ggatccctct aaaagcgggc atgacttcta gagtagtcca
      121 gggtttccga gggtttccgt cgacgatgtc agctcgtctc gagggcgtac taactgggcc
      181 gctacatgat atacgcacgg gtcctacaca acatactgca ggacaaactc ttcgcggtct
      241 ctgcatgcaa gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
      301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1141 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2461 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2521 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2581 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2641 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2701 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggacg
     2761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3181 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     3241 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     3301 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     3361 gacggccagt gaattg
//
