LOCUS       sequence-154442-        2529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTIM in vitro transcription backbone plasmid with two multiple 
            cloning sites and two Rho-independent transcriptional terminators..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2529)
  AUTHORS   Tapscott T, Kim JS, Crawford MA, Fitzsimmons L, Liu L, Jones-Carson 
            J, Vazquez-Torres A
  TITLE     Guanosine tetraphosphate relieves the negative regulation of 
            Salmonella pathogenicity island-2 gene transcription exerted by the 
            AT-rich ssrA discriminator region.
  JOURNAL   Sci Rep. 2018 Jun 21;8(1):9465. doi: 10.1038/s41598-018-27780-9.
  PUBMED    29930310
REFERENCE   2  (bases 1 to 2529)
  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: "Sci Rep."; 
            date: "2018-06-21"; volume: "8(1):9465. doi"; pages: " 
            10.1038/s41598-018-27780-9"
COMMENT     Sequence Label: pTIM
FEATURES             Location/Qualifiers
     source          1..2529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      255..341
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      433..460
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(737..753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(737..753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      801..832
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        833..935
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(936..1004,1005..1796)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1154..1173)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1868..1895
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1907..2494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1987..2006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcga ccgccggagg gttgcgtttg agacgggcga cagatcgaca ctgctcgatc
      181 cgctcgcacc atcgaattcg gtaccaagct tgctagccca tggctgcagg cgagagtagg
      241 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      301 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      361 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
      421 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttc
      481 ctgtcgtcga ctctagagag ctcatgcatg gatccgttaa ctctgggcgg ttctgataac
      541 gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt tttatggggg
      601 gagtttaggg agtggatcga cgagagcagc gcgactggat cagttctgga cgagcgagct
      661 gtcgtccgac ccgtgatctt acggcattat acgtatgatc ggtccacgat cagctagatt
      721 atctagtcag cttgatgtca tagctgtttc ctgaggctca atactgacca tttaaatcat
      781 acctgacctc catagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgatcggc
      841 acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg
      901 agttatcgag attttcagga gctaaggaag ctaaaatgag tattcaacat ttccgtgtcg
      961 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1021 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1081 tcaacagcgg taagatcctt gagagtttac gccccgaaga acgttttcca atgatgagca
     1141 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1201 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1261 agcatctcac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1321 ataacactgc ggccaactta cttctggcaa cgatcggagg accgaaggag ctaaccgctt
     1381 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1441 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1501 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1561 tggaggcgga taaagttgca ggatcacttc tgcgctcggc cctcccggct ggctggttta
     1621 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1681 cagatggtaa gccctcccgc atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1741 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaatgag
     1801 ggcccaaatg taatcacctg gctcaccttc gggtgggcct ttcttgagga cctaaatgta
     1861 atcacctggc tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc cataggctcc
     1921 gcccccctga cgagcatcac aaaaatcgat gctcaagtca gaggtggcga aacccgacag
     1981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttacctcg gaaaaagagt
     2401 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2461 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgattt tctaccgaag
     2521 aaaggccca
//
