LOCUS       sequence-11473-e        6195 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6195)
  AUTHORS   Yan M, Dai T, Deak JC, Kyriakis JM, Zon LI, Woodgett JR, Templeton 
            DJ
  TITLE     Activation of stress-activated protein kinase by MEKK1 
            phosphorylation of its activator SEK1.
  JOURNAL   Nature.   . 372(6508):798-800.
  PUBMED    7997270
REFERENCE   2  (bases 1 to 6195)
  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: "Nature.   .
            372(6508):798-800."
COMMENT     Sequence Label: HA SEK AL pcDNA3.1 zeo (+) - SEK1 (A-L)
FEATURES             Location/Qualifiers
     source          1..6195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     933..949
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     934..950
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             998..1024
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2141..2157)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2141..2157)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2202..2219)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2208..2432
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2478..2906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2565..2584)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2775..2796
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2915..2935)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2920..3250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3101..3236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3163..3182
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3298..3345
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3364..3738
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3868..3989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3905..3924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3959..3978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4038..4054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4038..4054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4051..4073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4062..4078
                     /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(4086..4092,4093..4110,4111..4116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4131..4152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4269..4286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4440..5028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4520..4539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5199..5990,5991..6059))
                     /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     5822..5841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6060..6164)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga
      961 tatcgaattc ctgcagcccc catctagagc caccatgtac ccatacgatg ttccagatta
     1021 cgctggaatt cgaaccatgg agggtaagcg caaacgactg aagttgaatt ttgcaaatcc
     1081 acctgtcaaa tcgacagcac ggtttaccct gaatcctaat actacaggag tccagaaccc
     1141 acacatagag agactgagaa cacacagcat tgagtcatca ggaaaactga agatctcccc
     1201 tgaacaacac tgggatttca ctgcagagga cttgaaagac cttggagaaa ttggacgagg
     1261 agcttatggt tctgtcaaca aaaggtccac aaacccaagt gggcagataa tggcagttaa
     1321 aagaattcgg tcaactgtgg atgaaaaaga acaaaaacaa cttctcatgg atttggatgt
     1381 agtaatgcgg agtagtgatt gcccatacat tgttcagttc tatggtgcac tcttcagaga
     1441 gggcgactgt tggatctgta tggagctcat gtctacctcg ttcgataagt tttacaaata
     1501 tgtatatagt gtgttagatg acgttattcc ggaagagatc ttaggcaaaa tcactttagc
     1561 aactgtgaaa gcactaaacc acttaaagaa acttgaaatt attcacagag acatcaaacc
     1621 ttccaatatt cttctggaca gaagtggaaa tataaagctc tgtgatttcg gcatcagtgg
     1681 acagcttgtg gacgctattg ccaagcttag agatgctggg tgtaggccgt atatggcacc
     1741 tgaaagaata gacccaagtg catcaagaca agggtatgat gtccgctctg atgtctggag
     1801 tttggggatc acattgtacg agttggccac acgatttcct tatccaaagt ggaatagtgt
     1861 atttgatcag ctaacacaag tggtgaaagg agaccctccg cagctgagta attctgaaga
     1921 aaggagttct ccccccagtt tcatcaactt tgtcaacttg tgccttacga aggatgaatc
     1981 caaaaggcca aagtataaag agcttctgaa acatcccttt attttgatgt atgaagaacg
     2041 tactgtagag gtcgcatgct atgtttgtaa aatcctggat cagatgccag ccactcccag
     2101 ctcgcccatg tatgttgact gaggatcctc gagtgggggg gatccactag ttctagagcg
     2161 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2221 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2281 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2341 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2401 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2461 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2521 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2581 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     2641 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2701 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2761 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2821 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     2881 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2941 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     3001 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3061 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3181 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3241 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     3301 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     3361 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     3421 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     3481 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     3541 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     3601 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     3661 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     3721 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
     3781 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3841 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     3901 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3961 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4021 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4201 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4621 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4861 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5161 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5221 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5281 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5341 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5401 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5461 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5521 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5581 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5641 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5701 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5761 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5821 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5881 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5941 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6001 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6061 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6121 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6181 gtgccacctg acgtc
//
