LOCUS       sequence-95180-e        3196 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Splitposon in KanR ColE1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3196)
  AUTHORS   Segall-Shapiro TH, Meyer AJ, Ellington AD, Sontag ED, Voigt CA
  TITLE     A 'resource allocator' for transcription based on a highly 
            fragmented T7 RNA polymerase.
  JOURNAL   Mol Syst Biol. 2014 Jul 30;10(7):742.
  PUBMED    25080493
REFERENCE   2  (bases 1 to 3196)
  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: "Mol Syst 
            Biol."; date: "2014-07-30"; volume: "10(7)"; pages: "742"
COMMENT     Sequence Label: pTHSSd_1
FEATURES             Location/Qualifiers
     source          1..3196
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(144..163)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             155..970
                     /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
                     ADRYQDLAILWNCLGEFSPSLQERLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(228..247)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(1275..1304)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1351..1372
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1407..2489)
                     /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
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2451..2470
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2490..2567)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        join(3086..3091,3092..3107,3108..3114)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3120..3139
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
ORIGIN
        1 tgattgattg aacgaaaaac gcgaaagcgt ttcacgataa atgcgaaaac ggatcctgtc
       61 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
      121 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
      181 ttgttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
      241 tcgcgataat gtcgggcaat caggtgcgac aatctatcgg ttgtatggga agcccgatgc
      301 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
      361 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
      421 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
      481 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
      541 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
      601 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
      661 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagctgt tgccattctc
      721 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
      781 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
      841 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacaggaac ggctttttca
      901 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
      961 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     1021 acgggacggc ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     1081 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     1141 tccacctaca acaactccgg caaaaaaacg ggcaaggtgt caccaccctg ccctttttct
     1201 ttaaaaccga aaagattact tcgcgtttgc cacctgacgt ctaagaaaag gaatattcag
     1261 caatttgccc gtgccgaaga aaggcccacc cgtgaaggtg agccagtgag ttgattgcta
     1321 cgtaattagt tagttagccc ttagtgactc gaattcgcgg ccgcttctag agcaattcgc
     1381 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1441 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     1501 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     1561 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     1621 ttgatggtgg ttgacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     1681 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     1741 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     1801 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     1861 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     1921 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     1981 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     2041 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     2101 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     2161 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     2221 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     2281 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     2341 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     2401 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     2461 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     2521 cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtcaac gtaaatgcat
     2581 gccgcttcgc cttcgcgcgc gaattggccg ccatgccggc gataatggcc tgcttctcgc
     2641 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     2701 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     2761 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     2821 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     2881 tcaagggcat cggtcgagat ccgggcttat cgactgcacg gtgcaccaat gcttctggcg
     2941 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     3001 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     3061 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     3121 attgtgagcg ctcacaatta ggatccgttt tcgcatttat cgtgaaacgc tttcgcattt
     3181 tgagtgaggt atatga
//
