LOCUS       sequence-242279-        3612 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 3612)
  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 3612)
  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_Gli3_Anti-Sense
FEATURES             Location/Qualifiers
     source          1..3612
                     /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(join(1140..1146,1147..1164,1165..1170))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1185..1206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1326..1343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1497..2085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1577..1596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2256..3047,3048..3116))
                     /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     2879..2898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3117..3221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3289..3307
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taatacgact cactataggg agagcggccg ccagatcttc cggatggctc gagtttttca
       61 gcaagatgtc tctcctgttg agtgtgttca gcactgtgaa gtctacacct gacaggtcac
      121 ttctgctcgg gaggagggtc ccgggcccac ccgagctata gttgttattg gactgtgtgc
      181 catttcctat gagaggagat accgcagggg ctttggaggg tagaataggg ttcagtcttg
      241 gaaacattcc attcacttgt tttagccttt caagtttgat gtgctccatc catttggtcc
      301 ctgccgggtt tctcctggct tgcaaagcaa gggctgtggt tgccgtggaa atggtcgagt
      361 ccatgattgg ggtttcatcg atggcactga ggtctgctac actacctcca tcagtcagag
      421 gaagctcgag cccactgttg gaatagttgc tgatggggga ctgttggctg ctgcatgaag
      481 actgaccacc agggcttggc tgagatgtca tgggcttctc agccttgaca gtcttcacct
      541 ggaggcactc ttcccgcttt gaggtagtgt tgctcagctc cttctgctca ccaaatgctc
      601 cctgagttgg ccggccaggg gacctggact gtgaatggct gccggaatct ctcgggggtg
      661 gaggccgagg gtgcatgtcc ccacgctgct tcttggtaac atgagatctt tctagaagat
      721 ctcctacaat attctcagct gccatggaaa atcgatgttc ttcttttatt ctctcaagat
      781 tttcaggctg tatattaaaa cttatattaa gaactatgct aaccacctca tcaggaaccg
      841 ttgtaggtgg cgtgggtttt cttggcaatc gactctcatg aaaactacga gctaaatatt
      901 caatatgttc ctcttgacca actttattct gcattttttt tgaacgaggt ttagagcaag
      961 cttcaggaaa ctgagacagg aattttatta aaaatttaaa ttttgaagaa agttcagggt
     1021 taatagcatc cattttttgc tttgcaagtt cctcagcatt cttaacaaaa gacgtctctt
     1081 ttgacatgtt taaagtttaa acctcctgtg tgaaattatt atccgctcat aattccacac
     1141 attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1201 acattaattg cgttgcgctc actgccaatt gctttccagt cgggaaacct gtcgtgccag
     1261 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1921 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2161 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2221 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2281 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2341 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2401 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2461 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2521 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2581 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2641 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2701 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2761 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2821 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2881 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2941 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3001 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3061 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3241 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     3301 acgaggccgc ccctgcagcc gaattatatt atttttgcca aataattttt aacaaaagct
     3361 ctgaagtctt cttcatttaa attcttagat gatacttcat ctggaaaatt gtcccaatta
     3421 gtagcatcac gctgtgagta agttctaaac cattttttta ttgttgtatt atctctaatc
     3481 ttactactcg atgagttttc ggtattatct ctatttttaa cttggagcag gttccattca
     3541 ttgttttttt catcatagtg aataaaatca actgctttaa cacttgtgcc tgaacaccat
     3601 atccatccgg cg
//
