LOCUS       sequence-248999-        5889 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 5889)
  AUTHORS   Horb ME, Thomsen GH
  TITLE     A vegetally localized T-box transcription factor in Xenopus eggs 
            specifies mesoderm and endoderm and is essential for embryonic 
            mesoderm formation.
  JOURNAL   Development. 1997 May;124(9):1689-98.
  PUBMED    9165117
REFERENCE   2  (bases 1 to 5889)
  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: 
            "Development."; date: "1997-05"; volume: "124(9)"; pages: "1689-98"
COMMENT     Sequence Label: pCS2-BRAT-EnR
FEATURES             Location/Qualifiers
     source          1..5889
                     /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    1941..2075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1965..1984)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2019..2038
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2188..2208)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2188..2206)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2227..2243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2227..2243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2240..2262)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2251..2267
                     /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(2275..2281,2282..2299,2300..2305))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2320..2341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2458..2475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2629..3217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2709..2728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3388..4179,4180..4248))
                     /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     4011..4030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4249..4353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4379..4834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4516..4537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4728..4747
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4905..5889
                     /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 aggtcgacac tagtggatcc
      121 aaagaattcg gcacgagcac ctgcagcaga gtttttatat tgagcattca cctgcagggg
      181 cccatgtgat agcctatagc tacagtgctg gaatgagaaa ctgctgtcgg gaatgcggcc
      241 tctctgcggg gcatctggaa ccagaggctt cttctaactg tgcatcagat gtaaagtctt
      301 ccccagacat ggacagtgtc tccagccaag actccctcta cctgcccaac actgttggtg
      361 cctccctgga agaccaagat ctatggtccc agttccacca ggaggggaca gagatgatca
      421 tcaccaaatc tggaaggagg atgtttcctc agtgtaagat ccgtctcttc ggtcttcatc
      481 cttatgccaa gtacatgctg ctggtggact ttgttcctct ggacaacttt aggtataagt
      541 ggaataagaa ccagtgggaa gcagcaggta aagcagaacc tcacccaccc tgcaggacgt
      601 atgtccaccc agattcacct gctcctggtg cccactggat gaaggatccg atctgctttc
      661 agaagctcaa actcaccaac aacacattgg atcaacaagg ccatattatc ttgcattcaa
      721 tgcatcgcta caagcccagg ttccatgtag ttcagtctga tgacatgtac aattctccat
      781 ggggattggt acaagtgttt agcttcccag agacagagtt tacttcagtg actgcctacc
      841 agaatgaaaa gattactaaa ctgaaaatta atcacaaccc atttgctaaa ggattccggg
      901 agcaggaaag gagtcacaag agggatgatg ttttaaagat tctacaacaa agtcccagta
      961 aaaggcagaa gaggaagaag tgggaggaca gtcctgaaat tcaaggtatg gccctggagg
     1021 atcgctgcag cccacagtca gcgcccagcc ccattaccct acaaatgcag catcttcacc
     1081 accagcaaca gcagcagcag caacagcagc agcaaatgca gcacctccac cagctgcagc
     1141 aactgcagca gttgcaccaa cagcaactgg ccgccggtgt cttccaccat ccggcaatgg
     1201 ccttcgatgc cgctgcagcc gccgctgctg cagctgctgc tgcggccgcc cacgctcatg
     1261 ctgctgcact gcagcagcgc ctcagtggca gtggatcgcc cgcatcctgc tccacgcccg
     1321 cctcgtccac gccgctgacc atcaaggagg aggaaagcga ctccgtgatc ggtgacatga
     1381 gtttccacaa tcagacgcac accaccaacg aggaggagga ggcggaggag gatgacgaca
     1441 ttgatgtgga tgtggatgat acgtcggcgg gcggacgcct gccaccaccc gcccaccagc
     1501 agcagtcgac ggccaagccc tcgctggcct tttccatctc caacatcctg agcgatcgtt
     1561 tcggagatgt ccagaagcct ggcaagtcga ttgagaacca ggccagcata ttccgcccct
     1621 tcgaggcgaa tcgctcccag actgccacgc cctccgcctt tacaagagtg gatctgctgg
     1681 agtttagccg gcaacagcaa gctgccgccg cagccgctac tgcggccatg atgctggaac
     1741 gggccaactt ccttaactgc ttcaatccgg ctgcctatcc caggatacac gaggaaatcg
     1801 tgcagagtcg cctgcgcagg agtgcagcca atgccgtcat cccgccgccc atgagctcca
     1861 agatgagcga tgccaatcca gagaaatctg ctctgggatc cctctcgagc ctctagaact
     1921 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
     1981 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2041 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2101 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc
     2161 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2221 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2281 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2341 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4381 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4441 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4501 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4561 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4621 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4681 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4741 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4801 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     4861 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca
     4921 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     4981 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     5041 atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg
     5101 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     5161 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     5221 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     5281 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     5341 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     5401 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     5461 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     5521 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     5581 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     5641 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     5701 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     5761 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     5821 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     5881 ttagggaac
//
