LOCUS       sequence-237444-        3613 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 3613)
  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 3613)
  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_Sense
FEATURES             Location/Qualifiers
     source          1..3613
                     /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(1410..1432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1421..1437
                     /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(1445..1451,1452..1469,1470..1475))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1490..1511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1631..1648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1802..2390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1882..1901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2561..3352,3353..3421))
                     /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     3184..3203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3422..3526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3594..3612
                     /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 tctcatgtta ccaagaagca gcgtggggac atgcaccctc ggcctccacc
      421 cccgagagat tccggcagcc attcacagtc caggtcccct ggccggccaa ctcagggagc
      481 atttggtgag cagaaggagc tgagcaacac tacctcaaag cgggaagagt gcctccaggt
      541 gaagactgtc aaggctgaga agcccatgac atctcagcca agccctggtg gtcagtcttc
      601 atgcagcagc caacagtccc ccatcagcaa ctattccaac agtgggctcg agcttcctct
      661 gactgatgga ggtagtgtag cagacctcag tgccatcgat gaaaccccaa tcatggactc
      721 gaccatttcc acggcaacca cagcccttgc tttgcaagcc aggagaaacc cggcagggac
      781 caaatggatg gagcacatca aacttgaaag gctaaaacaa gtgaatggaa tgtttccaag
      841 actgaaccct attctaccct ccaaagcccc tgcggtatct cctctcatag gaaatggcac
      901 acagtccaat aacaactata gctcgggtgg gcccgggacc ctcctcccga gcagaagtga
      961 cctgtcaggt gtagacttca cagtgctgaa cacactcaac aggagagaca atctttctag
     1021 aagatctcct acaatattct cagctgccat ggaaaatcga tgttcttctt ttattctctc
     1081 aagattttca ggctgtatat taaaacttat attaagaact atgctaacca cctcatcagg
     1141 aaccgttgta ggtggcgtgg gttttcttgg caatcgactc tcatgaaaac tacgagctaa
     1201 atattcaata tgttcctctt gaccaacttt attctgcatt ttttttgaac gaggtttaga
     1261 gcaagcttca ggaaactgag acaggaattt tattaaaaat ttaaattttg aagaaagttc
     1321 agggttaata gcatccattt tttgctttgc aagttcctca gcattcttaa caaaagacgt
     1381 ctcttttgac atgtttaaag tttaaacctc ctgtgtgaaa ttgttatccg ctcacaattc
     1441 cacacattat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1501 aactcacatt aattgcgttg cgctcactgc caattgcttt ccagtcggga aacctgtcgt
     1561 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1621 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1681 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1741 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2221 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     2281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2641 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2701 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2761 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2821 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2881 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2941 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3001 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3061 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3121 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3181 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3241 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3301 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3361 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3421 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3541 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3601 gtatcacgag gcc
//
