LOCUS       sequence-342226-        7247 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 7247)
  AUTHORS   Robinson MJ, Stippec SA, Goldsmith E, White MA, Cobb MH
  TITLE     A constitutively active and nuclear form of the MAP kinase ERK2 is 
            sufficient for neurite outgrowth and cell transformation.
  JOURNAL   Curr Biol. 1998 Oct 22;8(21):1141-50.
  PUBMED    9799732
REFERENCE   2  (bases 1 to 7247)
  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: "Curr 
            Biol."; date: "1998-10-22"; volume: "8(21)"; pages: "1141-50"
COMMENT     Sequence Label: pCMV-myc-ERK2-L4A-MEK1_fusion
FEATURES             Location/Qualifiers
     source          1..7247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..94
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     86..103
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     87..103
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        264..643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        644..847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     798..818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             876..905
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      925..934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1962..1997
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     polyA_signal    3534..4156
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3733..3752)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4180..4200)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4185..4514
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4365..4500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4552..4571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4553..4571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4585..4601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4585..4601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4598..4620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4609..4625
                     /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(4633..4639,4640..4657,4658..4663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4678..4699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4816..4833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4987..5575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5067..5086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5746..6537,6538..6606))
                     /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     6369..6388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6607..6711)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6738..7193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6825..6844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7035..7056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
       61 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac
      121 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
      181 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
      241 atgtccaata tgaccgccat gttgacattg attattgact agttattaat agtaatcaat
      301 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      361 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      421 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      481 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
      541 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
      601 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      661 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      721 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      781 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      841 cagagctcgt ttagtgaacc gtcagaattg ccatggagca gaagctgatc agcgaggagg
      901 acctgaacga attcagatct ggtaccatgg cggcggcggc ggcgggcccg gagatggtcc
      961 gcgggcaggt gttcgacgtg gggccgcgct acactaatct ctcgtacatc ggagaaggcg
     1021 cctacggcat ggtttgttct gcttatgata atctcaacaa agttcgagtt gctatcaaga
     1081 aaatcagtcc ttttgagcac cagacctact gtcagagaac cctgagagag ataaaaatcc
     1141 tactgcgctt cagacatgag aacatcatcg gcatcaatga catcatccgg gcaccaacca
     1201 ttgagcagat gaaagatgta tatatagtac aggacctcat ggagacagat ctttacaagc
     1261 tcttgaagac acagcacctc agcaatgatc atatctgcta ttttctttat cagatcctga
     1321 gaggattaaa gtatatacat tcagctaatg ttctgcaccg tgacctcaag ccttccaacc
     1381 tcctgctgaa caccacttgt gatctcaaga tctgtgactt tggccttgcc cgtgttgcag
     1441 atccagacca tgatcataca gggttcttga cagagtatgt agccacgcgt tggtacagag
     1501 ctccagaaat tatgttgaat tccaagggtt ataccaagtc cattgatatt tggtctgtgg
     1561 gctgcatcct ggcagagatg ctatccaaca ggcctatctt tccaggaaag cattaccttg
     1621 accagctgaa tcacatcctg ggtattcttg gatctccatc acaggaagat ctgaattgta
     1681 taataaattt aaaagctaga aactatttgc tttctctccc gcacaaaaat aaggtgccgt
     1741 ggaacagatt gttcccaaac gctgactcca aagctctgga tttactggat aaaatgttga
     1801 catttaaccc tcacaagagg attgaagttg aacaggctct ggcccacccg tacctggagc
     1861 agtattatga cccaagtgat gagcccattg ctgaagcacc attcaagttt gacatggagc
     1921 tggacgactt acctaaggag aagctcaaag aactcatttt tgaagagact gctcgattcc
     1981 agccaggata cagatctgaa ggcgagggtg aaggcgaggg tgaaagcgga tccggcggag
     2041 ttggaagcgc gttacccggg tccaaaatgc ccaagaagaa gccgacgccc atccagctga
     2101 acccggcccc cgacggctct gcagttaacg ggaccagctc tgcggagacc aacttggagg
     2161 ccgcgcagaa gaaggcggag gaggcagagg ctgatgagca gcagcgaaag cgccttgagg
     2221 cctttcttac ccagaagcag aaggtgggag aactgaagga tgacgacttt gagaagatca
     2281 gtgagctggg ggctggcaat ggcggtgtgg tgttcaaggt ctcccacaag ccttctggcc
     2341 tggtcatggc cagaaagcta attcatctgg agatcaaacc cgcaatccgg aaccagatca
     2401 taagggagct gcaggttctg catgagtgca actctccgta catcgtgggc ttctatggtg
     2461 cgttctacag cgatggcgag atcagtatct gcatggagca catggatgga ggttctctgg
     2521 atcaagtcct gaagaaagct ggaagaattc ctgaacaaat tttaggaaaa gttagcattg
     2581 ctgtaataaa aggcctgaca tatctgaggg agaagcacaa gatcatgcac agagatgtca
     2641 agccctccaa catcctagtc aactcccgtg gggagatcaa gctctgtgac tttggggtca
     2701 gcgggcagct catcgactcc atggccaact ccttcgtggg cacaaggtcc tacatgtcgc
     2761 cagaaagact ccaggggact cattactctg tgcagtcaga catctggagc atgggactgt
     2821 ctctggtaga gatggcggtt gggaggtatc ccatccctcc tccagatgcc aaggagctgg
     2881 agctgatgtt tgggtgccag gtggaaggag atgcggctga gaccccaccc aggccaagga
     2941 cccccgggag gccccttagc tcatacggaa tggacagccg acctcccatg gcaatttttg
     3001 agttgttgga ttacatagtc aacgagcctc ctccaaaact gcccagtgga gtgttcagtc
     3061 tggaatttca agattttgtg aataaatgct taataaaaaa ccccgcagag agagcagatt
     3121 tgaagcaact catggttcat gcttttatca agagatctga tgctgaggaa gtggattttg
     3181 caggttggct ctgctccacc atcggcctta accagcccag cacaccaacc catgctgctg
     3241 gcgtctaagt gtttgggaag caacaaagag cgagtcccct gcccggtggt ttgccatgtc
     3301 gcttttgggc ctccttccca tgcctgtctc tgttcagatg tgcatttcac ctgtgacaaa
     3361 ggatgaagaa cacagcatgt gccaagattc tactcttgtc atttttaata ttactgtctt
     3421 tattcttatt actattattg ttcccctaag tggattggct ttgtgcttgg ggctatttgt
     3481 gtgtatccga attcgaagct tgcatgcctg caggtcgact ctagagggtc cccgggtggc
     3541 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     3601 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     3661 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     3721 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     3781 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     3841 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     3901 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     3961 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa
     4021 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac
     4081 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt
     4141 agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt
     4201 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4261 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4321 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4381 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4441 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4501 taggcttttg caaaaagctc ctcgaggaac tgaaaaacca gaaagttaat tccctatagt
     4561 gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     4621 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4681 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4741 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6721 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6961 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7021 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7081 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7141 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg
     7201 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctc
//
