LOCUS       sequence-242277-        4891 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 4891)
  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 4891)
  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_Nkx3-2_Anti-Sense
FEATURES             Location/Qualifiers
     source          1..4891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(2400..2406,2407..2424,2425..2430))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2445..2466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2586..2603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2757..3345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2837..2856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3516..4307,4308..4376))
                     /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     4139..4158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4377..4481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4549..4567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     join(4873..4891,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4873..4891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagagcggcc gccagatctt ccggatggct cgagtttttc agcaagatgc gcttctttcg
       61 cggtttaggg gctgcgggct cttcctcctc ctccacgccg cccgcctgcc cgccgctagc
      121 cccgctgccc gcggcgccgc gcaacccagg cacgcgtgca cctccagggc taacgctgtc
      181 atcctcgcct ctcgggctgt ggtcgcctga aacgctggct gacatctcgc tgtcgctgcg
      241 gaccggggct tcctcctcca ggtccttggc ggcgtgcagc tcgcagcctg gctggtccag
      301 gcccagggtc accctggcgc ggccggttcc actggctccc gggacatcgg cgcagcgcct
      361 cctgccctcg ttctcttcgc tcagggctga gtctgagtcc caacctcccg gactctctgc
      421 gctctgcccc acggctgttc tggttcgggc gggagatgcc agaagagagt cctcggcgcc
      481 ccccagcgca ccggcctcgg tctccccgaa gatccgccag cagcagacgg cgggcgccgc
      541 ggtcaccgcc acctctgtgc cccctggtgc cgggcgcccc tccggcgtag ccaacccgcc
      601 gcgctcctct ttcttgttga ggatcgcatc tttctagaag atctcctaca atattctcag
      661 ctgccatgga aaatcgatgt tcttctctga gagatcccct cataatttcc ccaaagcgta
      721 accatgtgtg aataaatttt gagctagtag ggttgcagcc acgagtaagt cttcccttgt
      781 tattgtgtag ccagaatgcc gcaaaacttc catgcctaag cgaactgttg agagtacgtt
      841 tcgatttctg actgtgttag cctggaagtg cttgtcccaa ccttgtttct gagcatgaac
      901 gcccgcaagc caacatgtta gttgaagcat cagggcgatt agcagcatga tatcaaaacg
      961 ctctgagctg ctcgttcggc tatggcgtag gcctagtccg taggcaggac ttttcaagtc
     1021 tcggaaggtt tcttcaatct gcattcgctt cgaatagata ttaacaagtt gtttgggtgt
     1081 tcgaatttca acaggtaagt tagttgctag aacccatggc tcctttgccg acgctgagta
     1141 gattttaggt gacgggtggt gacaatgagt ccgtgtcgag cgctgatttt ttcggccttt
     1201 agagcgagat ttatacaata gaatttggca tgagattgga ttgcttttag tcagcctctt
     1261 atagcctaaa gtctttgagt gactagatga catatcatgt aagttgctga taggtttcca
     1321 gttttccgct cctaggtctg catattgtac ttttcctctt actcgactta accagtacca
     1381 acccagcttc tcaacggatt tataccatgg cactttaaag ccagcatcac tgacaatgag
     1441 cggtgtggtg ttactcggta gaatgctcgc aaggtcggct agaaattggt catgagcttt
     1501 ctttgaacat tgctctgaaa gcgggaacgc tttctcataa agagtaacag aacgaccgtg
     1561 tagtgcgact gaagctcgca ataccataag tcgtttttgc tcacgaatat cagaccagtc
     1621 aacaagtaca atgggcatcg tattgcccga acagataaag ctagcatgcc aacggtatac
     1681 agcgagtcgc tctttgtgga ggtgacgatt acctaacaat cggtcgattc gtttgatgtt
     1741 atgttttgtt ctcgctttgg ttggcaggtt acggccaagt tcggtaagag tgagagtttt
     1801 acagtcaagt aatgcgtggc aagccaacgt taagctgttg agtcgtttta agtgtaattc
     1861 ggggcagaat tggtaaagag agtcgtgtaa aatatcgagt tcgcacatct tgttgtctga
     1921 ttattgattt ttcgcgaaac catttgatca tatgacaaga tgtgtatcca ccttaactta
     1981 atgattttta ccaaaatcat taggggattc atcagtgttc ttcttttatt ctctcaagat
     2041 tttcaggctg tatattaaaa cttatattaa gaactatgct aaccacctca tcaggaaccg
     2101 ttgtaggtgg cgtgggtttt cttggcaatc gactctcatg aaaactacga gctaaatatt
     2161 caatatgttc ctcttgacca actttattct gcattttttt tgaacgaggt ttagagcaag
     2221 cttcaggaaa ctgagacagg aattttatta aaaatttaaa ttttgaagaa agttcagggt
     2281 taatagcatc cattttttgc tttgcaagtt cctcagcatt cttaacaaaa gacgtctctt
     2341 ttgacatgtt taaagtttaa acctcctgtg tgaaattatt atccgctcat aattccacac
     2401 attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2461 acattaattg cgttgcgctc actgccaatt gctttccagt cgggaaacct gtcgtgccag
     2521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3181 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4501 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4561 acgaggccgc ccctgcagcc gaattatatt atttttgcca aataattttt aacaaaagct
     4621 ctgaagtctt cttcatttaa attcttagat gatacttcat ctggaaaatt gtcccaatta
     4681 gtagcatcac gctgtgagta agttctaaac cattttttta ttgttgtatt atctctaatc
     4741 ttactactcg atgagttttc ggtattatct ctatttttaa cttggagcag gttccattca
     4801 ttgttttttt catcatagtg aataaaatca actgctttaa cacttgtgcc tgaacaccat
     4861 atccatccgg cgtaatacga ctcactatag g
//
