LOCUS       sequence-237441-        3759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for anti-sense in situ probe in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3759)
  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 3759)
  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_Rab28_Anti-Sense
FEATURES             Location/Qualifiers
     source          1..3759
                     /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(1556..1578)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1567..1583
                     /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(1591..1597,1598..1615,1616..1621))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1636..1657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1777..1794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1948..2536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2028..2047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2707..3498,3499..3567))
                     /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     3330..3349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3568..3672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3740..3758
                     /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 tgatcatcaa agcttggagc agcttcatat ctggttaaat tttttatatc
      421 tgggaaacaa agaagtgaag agagaaggaa gcaacatggt tcttctggag tgaggaggca
      481 gtctcaatcc agagaatgga ctctccattc ggatttcaaa acgtgtggtt ccaaatcgca
      541 ctccttccgg tccagggagc tgcctgatcc ctcaggcaac taaaggatga aggagcagag
      601 caggaggaag tgtgagctgg ctacggagac agtcactgat aaaacacgtt cctaggcagc
      661 tggggcccaa ttcaggggaa gagcagccag aactatcaat acagtgtgtg ctcactgcac
      721 tgcacacatg gagctccgag gagggttaac agttcttgac aagggctcct ggttgtagtt
      781 tacaatatct gccttcacca ctctctgtga ctgttctatt tctgctttgt ttaatttgat
      841 tcccaggatt tctgctgcaa ctttctgaaa acacaggaag acagagtctc ctgtcttggc
      901 tgaaacaaag tggctactaa aaccattttc ctggcaaaat cgtaagtgct tatcagcttt
      961 tactgttcgc atatgctcca agtcaatttt attgcccact aaagcaacca ggggctgagt
     1021 ttctgactct tcacttactg tcttcaccac agaataccaa tcttccaaat tctcaaagct
     1081 ttggtagttt gtaatatcat ataccaagag gattccctgt gctccataga tgtatttatc
     1141 caacatcttg ccacctatct ttctagaaga tctcctacaa tattctcagc tgccatggaa
     1201 aatcgatgtt cttcttttat tctctcaaga ttttcaggct gtatattaaa acttatatta
     1261 agaactatgc taaccacctc atcaggaacc gttgtaggtg gcgtgggttt tcttggcaat
     1321 cgactctcat gaaaactacg agctaaatat tcaatatgtt cctcttgacc aactttattc
     1381 tgcatttttt ttgaacgagg tttagagcaa gcttcaggaa actgagacag gaattttatt
     1441 aaaaatttaa attttgaaga aagttcaggg ttaatagcat ccattttttg ctttgcaagt
     1501 tcctcagcat tcttaacaaa agacgtctct tttgacatgt ttaaagttta aacctcctgt
     1561 gtgaaattgt tatccgctca caattccaca cattatacga gccggaagca taaagtgtaa
     1621 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgccaat
     1681 tgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1741 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1801 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1861 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1921 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1981 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2281 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2341 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     2401 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2461 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2761 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2821 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2881 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2941 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3001 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3061 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3121 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3181 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3241 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3301 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3361 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3421 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3481 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3541 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3601 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3661 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3721 catgacatta acctataaaa ataggcgtat cacgaggcc
//
