LOCUS       sequence-249058-        4982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4982)
  AUTHORS   Nishimatsu S, Thomsen GH
  TITLE     Ventral mesoderm induction and patterning by bone morphogenetic 
            protein heterodimers in Xenopus embryos.
  JOURNAL   Mech Dev. 1998 Jun;74(1-2):75-88.
  PUBMED    9651485
REFERENCE   2  (bases 1 to 4982)
  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: "Mech Dev.";
            date: "1998-06"; volume: "74(1-2)"; pages: "75-88"
COMMENT     Sequence Label: p64T-XBMP2
FEATURES             Location/Qualifiers
     source          1..4982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2069..2085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2069..2085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2082..2104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2093..2109
                     /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(2117..2123,2124..2141,2142..2147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2162..2183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2300..2317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2471..3059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2551..2570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3230..4021,4022..4090))
                     /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     3853..3872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4091..4195)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4263..4281
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4319..4341)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4441..4460
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4674..4693
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atttaggtga cactatagaa tacaagcttg ggctgcaggt caattccggc tcattgccga
       61 cggacacggc gatcagtcgc agcgactatg agcagaaagc gaaggactgg gctgcgagca
      121 gacgcacaga ctcgccctgg gaaggagacg cgtggagtaa aaagactcga tcgagtgatg
      181 tgggaaaaac acacggacag cagaaaacca tcatcgagac acaaagtcgg cggcgcacag
      241 gttgacaatg gtcgctggga tccaccctct gctcctgctg ctgttttacc agctcttgct
      301 gagtggctgc actggactga tacccgagga aggcaagcgc aagtatacgg aatcgggtcg
      361 ctcgtctccc cagcagtccc agagggtcct caaccagttc gagctccggc tgctcagtat
      421 gtttggcttg aagaggcgtc cgacgcctgg caaaaatgtt gtgatcccac cctacatgct
      481 ggacttgtac cacctgcact tggctcagtt ggctgctgat gaaggcactt ctgccatgga
      541 cttccagatg gagcgggcag ccagccgagc aaatacagtg aggagtttcc accatgaaga
      601 atccatggaa gaaattcctg agtctcgtga gaaaacaatc caacgattct tcttcaacct
      661 ttcttcaatt ccaaatgagg agctggtcac ttctgccgag ctgcggattt ttcgagagca
      721 ggtccaagag ccctttgaaa gtgacagcag caaattgcat cggattaata tttacgacat
      781 tgtcaagcca gcggcggctg cctcccgggg ccctgttgtg agactattgg acaccagact
      841 ggtacatcat aatgaaagca aatgggaaag ttttgatgta acgccggcaa ttgcgcggtg
      901 gattgcacat aaacagccta accatgggtt tgttgttgaa gtgactcact tggacaatga
      961 caaaaatgtg cctaagaagc atgtgaggat tagtaggtct ttaaccccgg ataaagataa
     1021 ctggcctcag atacggccat tgttggtaac ttttagccat gatggtaaag gacatgctct
     1081 tcacaaaaga caaaagcgcc aagctaggca caaacaacgt aaacgcctta aatcgagctg
     1141 caggaggcat ccgttgtacg tagatttcag cgacgttggt tggaatgact ggattgttgc
     1201 cccacctggg tatcatgcct tttactgcca cggggaatgt ccttttccac tggcagacca
     1261 tttaaactct acaaaccatg caatcgtaca aactttggtg aactctgtta acacaaacat
     1321 ccccaaagct tgctgcgtcc ccacggaact cagtgccata tccatgctct atcttgatga
     1381 gaatgaaaaa gtagtattaa aaaattatca agacatggtc gtggaggggt gcggatgccg
     1441 ttaggcagtt acgcgcaagc cagagacaag aaagatgaca ctttaatatt tccttttgga
     1501 gactatattt atgctttgaa aaatgatgaa acaattattt tgaaaatata tttatgtcta
     1561 cacggaggtt gggaagcaaa tattttaatc agagaaatat tccttttttt agttgtacat
     1621 ttttataagg gtttgtaccc agcacatgaa gtataatggt cagattccta ttttgtattt
     1681 atttaccatt ataaccactt ttttagggaa aaaatagctg ttttgtattt atatgtaatc
     1741 aacagagaaa atatagggtt tgtaaaaatg ttactgaaag cagattcatt ggttttcttt
     1801 ttttaagtta tacacagctg gttatatggc aagtgttata ttttctacaa agctaatttt
     1861 aaggtcagtt gttataaaaa tgataatgtg ttggttcatt tttatatcct tcatattgtg
     1921 ccattaacat gcatttttta aaatgtagga agtccagtgt gcattgcttt gtaaattcag
     1981 aattggaatt actctagcta gaggatcccc gggcgagctc ccaaaaaaaa aaaaaaaaaa
     2041 aaaaaaaaaa aaccgaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     2101 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2461 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2521 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2581 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2641 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2701 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2761 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2821 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2881 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2941 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3001 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3061 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3121 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3181 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3241 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3301 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3361 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3421 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3481 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3541 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3601 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3661 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3721 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3781 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3841 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3901 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3961 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4021 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4081 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4141 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4201 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4261 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4321 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4381 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4441 cggcatcaga gcagattgta ctgagagtgc accattcgac gctctccctt atgcgactcc
     4501 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     4561 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     4621 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     4681 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     4741 ccggcgtaga ggatctggct agcgatgacc ctgctgattg gttcgctgac catttccggg
     4801 tgcgggacgg cgttaccaga aactcagaag gttcgtccaa ccaaaccgac tctgacggca
     4861 gtttacgaga gagatgatag ggtctgcttc agtaagccag atgctacaca attaggcttg
     4921 tacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     4981 cg
//
