LOCUS       sequence-243532-        3553 bp    DNA     circular SYN 15-APR-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 3553)
  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 3553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_Sense
FEATURES             Location/Qualifiers
     source          1..3553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1062..1068,1069..1086,1087..1092))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1107..1128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1248..1265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1419..2007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1499..1518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2178..2969,2970..3038))
                     /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     2801..2820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3039..3143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3211..3229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     join(3535..3553,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3535..3553
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagagcggcc gccagatctt ccggatggct cgagtttttc agcaagatgc gatcctcaac
       61 aagaaagagg agcgcggcgg gttggctacg ccggaggggc gcccggcacc agggggcaca
      121 gaggtggcgg tgaccgcggc gcccgccgtc tgctgctggc ggatcttcgg ggagaccgag
      181 gccggtgcgc tggggggcgc cgaggactct cttctggcat ctcccgcccg aaccagaaca
      241 gccgtggggc agagcgcaga gagtccggga ggttgggact cagactcagc cctgagcgaa
      301 gagaacgagg gcaggaggcg ctgcgccgat gtcccgggag ccagtggaac cggccgcgcc
      361 agggtgaccc tgggcctgga ccagccaggc tgcgagctgc acgccgccaa ggacctggag
      421 gaggaagccc cggtccgcag cgacagcgag atgtcagcca gcgtttcagg cgaccacagc
      481 ccgagaggcg aggatgacag cgttagccct ggaggtgcac gcgtgcctgg gttgcgcggc
      541 gccgcgggca gcggggctag cggcgggcag gcgggcggcg tggaggagga ggaagagccc
      601 gcagccccta aaccgcgaaa gaagcgcatc tttctagaag atctcctaca atattctcag
      661 ctgccatgga aaatcgatgt tcttctttta ttctctcaag attttcaggc tgtatattaa
      721 aacttatatt aagaactatg ctaaccacct catcaggaac cgttgtaggt ggcgtgggtt
      781 ttcttggcaa tcgactctca tgaaaactac gagctaaata ttcaatatgt tcctcttgac
      841 caactttatt ctgcattttt tttgaacgag gtttagagca agcttcagga aactgagaca
      901 ggaattttat taaaaattta aattttgaag aaagttcagg gttaatagca tccatttttt
      961 gctttgcaag ttcctcagca ttcttaacaa aagacgtctc ttttgacatg tttaaagttt
     1021 aaacctcctg tgtgaaatta ttatccgctc ataattccac acattatacg agccggaagc
     1081 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1141 tcactgccaa ttgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1261 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1321 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1381 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1441 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1501 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1561 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1621 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1681 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1741 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1801 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1861 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1921 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2101 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3061 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3121 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3181 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc gcccctgcag
     3241 ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc ttcttcattt
     3301 aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc acgctgtgag
     3361 taagttctaa accatttttt tattgttgta ttatctctaa tcttactact cgatgagttt
     3421 tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt ttcatcatag
     3481 tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc ggcgtaatac
     3541 gactcactat agg
//
