LOCUS       sequence-248972-        5616 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 5616)
  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 5616)
  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: pCS2(+)XBMP7
FEATURES             Location/Qualifiers
     source          1..5616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1668..1802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1692..1711)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1746..1765
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1915..1935)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1915..1933)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1954..1970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1954..1970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1967..1989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1978..1994
                     /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(2002..2008,2009..2026,2027..2032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2047..2068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2185..2202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2356..2944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2436..2455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3115..3906,3907..3975))
                     /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     3738..3757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3976..4080)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4106..4561)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4243..4264)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4455..4474
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4632..5616
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggcctctcg aggaattccg
      121 atatggaatg taaaaatact ggtgaattat gggaagtccg acacagacca gtaacttcag
      181 catgttatct ttgacaaaat gaatgctttg acagtaaaga gaagattgcc tgtgctgctt
      241 tttctttttc acatttcact gagttccatc tcgtcaaata caatattgga gaatgatttc
      301 cactctagtt ttgtccagag aagactaaaa ggccacgaac gcagagagat tcaaaaagag
      361 atcttgacta ttttaggttt gcaacacaga ccaaggccat atttaccgga gaaaaagaag
      421 tctgcaccat tattcatgat ggatttatac aatgcagtaa atattgaaga gatgcatgct
      481 gaagatgttt cctacagcaa taagccgatc tccctaaatg aagctttttc actggccact
      541 gaccaagaga atggctttct tgcacatgcc gacacagtta tgagttttgc taatttagtt
      601 gacaatgaca acgaattgca taaaaactcc tatcgccaaa aattcaagtt tgatctaact
      661 gatatcccac ttggagatga actgacagcc gctgaatttc gaatttataa agattatgta
      721 caaaataacg agacatacca ggtcaccatc taccaggtgc ttaagaagca agccgacaaa
      781 gatccttatc ttttccaggt agactcaaga accatctggg gcacagaaaa gggatggctg
      841 acgtttgata ttactgcaac tggtaatcac tgggtgatga acccacatta caaccttgga
      901 ttgcagttat cagtagagag tatggatatg caaaatgtta atcccaggct tgtgggcctt
      961 gttggaaaga atggtcctca agacaaacag ccatttatgg tggcattctt taagacctca
     1021 gatatccatc tccgcagtgt tcgatctact agcaataagc actggaatca ggaaagagcc
     1081 aagacctaca aggagcaaga taatttacct ccagcaaata ttactgatgg catcatgccc
     1141 cctggaaaac gtcgtttttt aaagcaagct tgcaagaaac atgaactgtt tgtaagtttc
     1201 cgcgatcttg gttggcaaga ctggataatt gcacctgaag gatatgctgc ctactattgt
     1261 gatggagaat gtgctttccc acttaactct ttcatgaatg ccacaaacca tgccattgta
     1321 caaacgttgg tacatttcat taacccagag actgtcccta agccatgctg tgcaccaact
     1381 cagctcaatg gtatttctgt tttatacttt gatgacagtg ccaatgttat attaaagaaa
     1441 tacaaaaata tggtggttca agcctgtggt tgccattgac aatagcagtt attctgtttt
     1501 taacagtcat tttaatggta ttgtccttat gtttatttta aagtagagat acttgaccat
     1561 cacacttaaa aaaatgcatt gtacacctta acggatgaaa agattttgtt tttgcatgat
     1621 ttcggaattc ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1681 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1741 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1801 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1861 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1921 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1981 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     2041 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2101 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2161 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2221 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2281 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2341 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2821 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2881 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3061 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3121 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3181 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3241 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3301 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3361 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3421 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3481 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3541 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3601 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3661 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3721 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3781 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3841 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3901 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3961 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4021 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4081 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4141 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4201 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4261 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4321 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4381 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4441 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4501 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4561 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4621 ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata
     4681 tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata
     4741 tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata
     4801 tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga
     4861 ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg
     4921 cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc
     4981 caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca
     5041 tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata
     5101 tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat
     5161 atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca
     5221 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
     5281 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
     5341 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
     5401 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
     5461 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
     5521 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
     5581 tgggaggtct atataagcaa tgctcgttta gggaac
//
