LOCUS       sequence-242686-        5488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5488)
  AUTHORS   Creasy CL, Ambrose DM, Chernoff J
  TITLE     The Ste20-like protein kinase, Mst1, dimerizes and contains an 
            inhibitory domain.
  JOURNAL   J Biol Chem. 1996 Aug 30. 271(35):21049-53.
  PUBMED    8702870
REFERENCE   2  (bases 1 to 5488)
  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: "J Biol 
            Chem. 1996 Aug 30. 271(35):21049-53."
COMMENT     Sequence Label: pJ3H-Mst2 K56R
FEATURES             Location/Qualifiers
     source          1..5488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..54
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     RBS             723..731
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     intron          2161..2226
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2356..2376
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2801..2935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2838..2857)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2892..2911
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2966..3070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3071..3139,3140..3931)
                     /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     complement(3289..3308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4102..4690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4591..4610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4844..4861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4876..5016
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4942..4961)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5102..5124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5162..5182)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(5167..5488,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5347..5482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaag cttcgatcgt cgacatgtac ccatacgatg ttccagatta cgcttctaga
       61 ggatccgcca tggagcagcc gccggcgcct aagagtaaac taaaaaagct gagtgaagac
      121 agtttgacta agcagcctga agaagttttt gatgtattag agaagcttgg agaagggtct
      181 tatggaagtg tatttaaagc aatacacaag gaatccggtc aagttgtcgc aattagacaa
      241 gtacctgttg aatcagatct tcaggaaata atcaaagaaa tttccataat gcagcaatgt
      301 gacagcccat atgttgtaaa gtactatggc agttatttta agaatacaga cctctggatt
      361 gttatggagt actgtggcgc tggctctgtc tcagacataa ttagattacg aaacaagaca
      421 ttaatagaag atgaaattgc aaccattctt aaatctacat tgaaaggact agaatatttg
      481 cactttatga gaaaaataca cagagatata aaagctggaa atattctcct caatacagaa
      541 ggacatgcaa aattggcaga ttttggagtg gctggtcagt taacagatac aatggcaaaa
      601 cgcaatactg taataggaac tccattttgg atggctcctg aggtgattca agaaataggc
      661 tataactgtg tggccgacat ctggtccctt ggcattactt ctatagaaat ggctgaagga
      721 aaacctcctt atgctgatat acatccaatg agggctattt ttatgattcc cacaaatcca
      781 ccaccaacat tcagaaagcc agaactttgg tccgatgatt tcaccgattt tgttaaaaag
      841 tgtttggtga agaatcctga gcagagagct actgcaacac aacttttaca gcatcctttt
      901 atcaagaatg ccaaacctgt atcaatatta agagacctga tcacagaagc tatggagatc
      961 aaagctaaaa gacatgacga acagcaacga gaattggaag aggaagaaga aaattcggat
     1021 gaagatgagc tggattccca caccatggtg aagactagtg tggagagtgt gggcaccatg
     1081 cgggccacaa gcatgatgag tgaaggggcc cagaccatga ttgaacataa tagcacgatg
     1141 ttggaatccg acttggggac catggtgata aacagtgagg atgaggaaga agaagatgga
     1201 actatgaaaa gaaatgcaac ctcaccacaa gtacaaagac catctttcat ggactacttt
     1261 gataagcaag acttcaagaa taagagtcac gaaaactgta atcagaacat gcatgaaccc
     1321 ttccctatgt ccaaaaacgt ttttcctgat aactggaaag ttcctcaaga tggagacttt
     1381 gactttttga aaaatctaag tttagaagaa ctacagatgc ggttaaaagc actggacccc
     1441 atgatggaac gggagataga agaacttcgt cagagataca ctgcgaaaag acagcccatt
     1501 ctggatgcga tggatgcaaa gaaaagaagg cagcaaaact tttgagtcta atttcctctc
     1561 tgtttttaac tattctggag accaagaaac cactaggaat tgaaggaata tttggatatt
     1621 tttaatccta agattttgcc ctacaattag gcagaggtca aaaagtgaca atggtacatg
     1681 cccaggtaaa ttcccaaaag gcagaattga cagttgtatc tgctgtgcat tcactctaag
     1741 atgaggagaa caaaagaagt gtattctctt gttctgtcag ctgcatacca gtaataaaac
     1801 tgttatgaaa tggattttca aggtctctaa accttgaaaa tccaaagcta ttgttgcatt
     1861 gtacagcact gaagggcttt atgttacaat attctttatt cctatctagt atactaggct
     1921 atttattgta tccccttagg taaacttatt tatttatgct attttgcttt gtttcatttt
     1981 ttaaggacaa gatcaggata gctttggtga aggtagggtc atattaatat gatgataatg
     2041 tgcaaccaat ttatactttc tgcaggaatt catcgatggt accagatctt tgtgaaggaa
     2101 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     2161 gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta
     2221 ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc ctttaatgag
     2281 gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac tgctgactct
     2341 caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga ctttccttca
     2401 gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc ttgctttgct
     2461 atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga aaaatattct
     2521 gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt tcttactcca
     2581 cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac ctttagcttt
     2641 ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac tagagatcat
     2701 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2761 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2821 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2881 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctctgc
     2941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3061 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3121 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3181 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3241 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3301 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3361 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3421 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3481 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3541 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3601 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3661 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3721 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3781 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3841 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3901 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3961 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4021 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4081 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4141 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4201 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4261 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4321 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4381 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4441 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4501 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4561 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4621 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4681 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4741 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4801 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4861 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4921 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4981 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     5041 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5101 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     5161 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5221 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5341 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5401 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5461 tgaggaggct tttttggagg cctaggct
//
