LOCUS       sequence-288037-        5782 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5782)
  AUTHORS   Robinson FL, Whitehurst AW, Raman M, Cobb MH
  TITLE     Identification of novel point mutations in ERK2 that selectively 
            disrupt binding to MEK1.
  JOURNAL   J Biol Chem. 2002 Apr 26;277(17):14844-52. Epub 2002 Jan 31.
  PUBMED    11823456
REFERENCE   2  (bases 1 to 5782)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2002-04-26"; volume: "277(17)"; pages: "14844-52. 
            Epub 2002 Jan 31"
COMMENT     Sequence Label: p3xFlag-CMV7-Erk2
FEATURES             Location/Qualifiers
     source          1..5782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2065..2100
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     polyA_signal    2123..2745
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2322..2341)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2769..2789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2774..3103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2954..3089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3141..3160)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3142..3160)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3174..3190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3174..3190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3187..3209)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3198..3214
                     /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(3222..3228,3229..3246,3247..3252))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3267..3288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3405..3422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3576..4164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3656..3675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4335..5126,5127..5195))
                     /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     4958..4977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5196..5300)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5327..5755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5414..5433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5624..5645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcatcgata
     1021 gatcccatca tggcggcggc ggcggcgggc ccggagatgg tccgcgggca ggtgttcgac
     1081 gtggggccgc gctacactaa tctctcgtac atcggagaag gcgcctacgg catggtttgt
     1141 tctgcttatg ataatctcaa caaagttcga gttgctatca agaaaatcag tccttttgag
     1201 caccagacct actgtcagag aaccctgaga gagataaaaa tcctactgcg cttcagacat
     1261 gagaacatca tcggcatcaa tgacatcatc cgggcaccaa ccattgagca gatgaaagat
     1321 gtatatatag tacaggacct catggagaca gatctttaca agctcttgaa gacacagcac
     1381 ctcagcaatg atcatatctg ctattttctt tatcagatcc tgagaggatt aaagtatata
     1441 cattcagcta atgttctgca ccgtgacctc aagccttcca acctcctgct gaacaccact
     1501 tgtgatctca agatctgtga ctttggcctt gcccgtgttg cagatccaga ccatgatcat
     1561 acagggttct tgacagagta tgtagccacg cgttggtaca gagctccaga aattatgttg
     1621 aattccaagg gttataccaa gtccattgat atttggtctg tgggctgcat cctggcagag
     1681 atgctatcca acaggcctat cttcccagga aagcattacc ttgaccagct gaatcacatc
     1741 ctgggtattc ttggatctcc atcacaggaa gatctgaatt gtataataaa tttaaaagct
     1801 agaaactatt tgctttctct cccgcacaaa aataaggtgc cgtggaacag attgttccca
     1861 aacgctgact ccaaagctct ggatttactg gataaaatgt tgacatttaa ccctcacaag
     1921 aggattgaag ttgaacaggc tctggcccac ccgtacctgg agcagtatta tgacccaagt
     1981 gatgagccca ttgctgaagc accattcaag tttgacatgg agctggacga cttacctaag
     2041 gagaagctca aagaactcat ttttgaagag actgctcgat tccagccagg atacagatct
     2101 tgactcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
     2161 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2221 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     2281 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     2341 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     2401 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     2461 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     2521 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     2581 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
     2641 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
     2701 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
     2761 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2941 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3061 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact
     3121 gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag
     3181 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3241 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3301 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3361 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3421 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3481 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3541 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3601 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3661 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3721 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3781 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3841 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3901 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3961 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4021 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4081 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4141 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4201 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4261 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4321 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4381 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4441 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4501 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4561 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4621 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4681 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4741 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4801 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4861 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4921 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4981 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5041 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5101 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5161 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5221 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5281 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     5341 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5401 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5461 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5521 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5581 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5641 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5701 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5761 aaaatattaa cgcttacaat tt
//
