LOCUS       sequence-237440-        3847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for sense in situ probe in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3847)
  AUTHORS   Castro JP, Yancoskie MN, Marchini M, Belohlavy S, Hiramatsu L, Kucka
            M, Beluch WH, Naumann R, Skuplik I, Cobb J, Barton NH, Rolian C, 
            Chan YF
  TITLE     An integrative genomic analysis of the Longshanks selection 
            experiment for longer limbs in mice.
  JOURNAL   Elife. 2019 Jun 6;8:e42014. doi: 10.7554/eLife.42014.
  PUBMED    31169497
REFERENCE   2  (bases 1 to 3847)
  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: "2019-06-6"; volume: "8:e42014. doi"; pages: " 
            10.7554/eLife.42014"
COMMENT     Sequence Label: pJET1.2_Bod1l_Sense
FEATURES             Location/Qualifiers
     source          1..3847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1644..1666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1655..1671
                     /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(1679..1685,1686..1703,1704..1709))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1724..1745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1865..1882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2036..2624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2116..2135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2795..3586,3587..3655))
                     /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     3418..3437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3656..3760)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3828..3846
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tgatgccatg tcaatcttgg aaaccataac ttctcttaac caagaagcta
      421 acgctgccag agcttcaaca gaaatgtcaa atgccaaggt cagtgagaga acatccagaa
      481 aactctcatc tcagccaagc actgacgtta gcacagacaa ggagagaggc tcagaagatg
      541 cgactgagag agaaaaggct acctctgatt ctggaggaga tgggctagaa gctgctctca
      601 agtcagaaga acccagtgat ctcccttgtc cagtggaaga aactaaaaat catatgaaag
      661 agaataatag cttactgtta ctaagtaagg atgctcagca ggaaagcact gacccaaaaa
      721 ttaaatcaat ggacaaagga gaaaagaaac ccgatggtaa tgagaaagga gagagaaaga
      781 aagagaagaa ggaaaagact gaaaaaaaaa ttgatcactc taaaaggaat gaagatacac
      841 agaaagtgaa agatgagaga caagcaaagg acaaagaagt ggagagcaca aagcttcctt
      901 ctgaaaagag caacagcaga gccagggctg ctgaggggac aaaagaagat tgttctttgt
      961 tagattctga tgtggatggt ctcactgata tcacagtcag ctctgtccac accagtgacc
     1021 tttcgtcctt tgaagaagac acagaggagg aagttgtcgt gtctgagagc atggaagaag
     1081 gagagattac atcagaggat gaagagaaaa acaagcagaa taaagcaaaa gttcaacctg
     1141 gtgattctag tgatggaaaa gccagaggcg tacggcatgc ttatgttcac aaaccgtatc
     1201 tttactcaaa gtactacagt gattctgatg acgaactcac agtgatcttt ctagaagatc
     1261 tcctacaata ttctcagctg ccatggaaaa tcgatgttct tcttttattc tctcaagatt
     1321 ttcaggctgt atattaaaac ttatattaag aactatgcta accacctcat caggaaccgt
     1381 tgtaggtggc gtgggttttc ttggcaatcg actctcatga aaactacgag ctaaatattc
     1441 aatatgttcc tcttgaccaa ctttattctg catttttttt gaacgaggtt tagagcaagc
     1501 ttcaggaaac tgagacagga attttattaa aaatttaaat tttgaagaaa gttcagggtt
     1561 aatagcatcc attttttgct ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt
     1621 tgacatgttt aaagtttaaa cctcctgtgt gaaattgtta tccgctcaca attccacaca
     1681 ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1741 cattaattgc gttgcgctca ctgccaattg ctttccagtc gggaaacctg tcgtgccagc
     1801 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1861 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3781 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3841 cgaggcc
//
