LOCUS       sequence-311041-        5918 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 5918)
  AUTHORS   Yao Z, Flash I, Raviv Z, Yung Y, Asscher Y, Pleban S, Seger R
  TITLE     Non-regulated and stimulated mechanisms cooperate in the nuclear 
            accumulation of MEK1.
  JOURNAL   Oncogene. 2001 Nov 15. 20(52):7588-96.
  PUBMED    11753637
REFERENCE   2  (bases 1 to 5918)
  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: "Oncogene. 
            2001 Nov 15. 20(52):7588-96."
COMMENT     Sequence Label: MEK1-GFP
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      590..599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1799..1808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1805..1807,1808..1810,1811..2524)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1805..1807,1808..1810,1811..2521)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1850..1871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2111..2130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2458..2479
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2647..2768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2684..2703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2738..2757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2775..3230)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2912..2933)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3124..3143
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3257..3361
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3363..3720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3363..3554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3386..3406)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3571..3706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3755..4549
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3755..4549
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3809..3828)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4419..4438
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4737..4756)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4781..4828
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5157..5745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5646..5665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggactca gatctcgagc tcaagcttgg taccgagctc ggatccgccg ccaccatggc
      601 acccaagaag aagccgacgc ccatccagct gaacccggcc cccgacggct ctgcagttaa
      661 cgggaccagc tctgcggaga ccaacttgga ggccttgcag aagaagctgg aggagctaga
      721 gcttgatgag cagcagcgaa agcgccttga ggcctttctt acccagaagc agaaggtggg
      781 agaactgaag gatgacgact ttgagaagat cagtgagctg ggggctggca atggcggtgt
      841 ggtgttcaag gtctcccaca agccttctgg cctggtcatg gccagaaagc taattcatct
      901 ggagatcaaa cccgcaatcc ggaaccagat cataagggag ctgcaggttc tgcatgagtg
      961 caactctccg tacatcgtgg gcttctatgg tgcgttctac agcgatggcg agatcagtat
     1021 ctgcatggag cacatggatg gaggttctct ggatcaagtc ctgaagaaag ctggaagaat
     1081 tcctgaacaa attttaggaa aagttagcat tgctgtaata aaaggcctga catatctgag
     1141 ggagaagcac aagatcatgc acagagatgt caagccttcc aacatcctag tcaactcccg
     1201 tggggagatc aagctctgtg actttggggt cagcgggcag ctcatcgact ccatggccaa
     1261 ctccttcgtg ggcacaaggt cctacatgtc gccagaaaga ctccagggga ctcattactc
     1321 tgtgcagtca gacatctgga gcatgggact gtctctggta gagatggcgg ttgggaggta
     1381 tcccatccct cctccagatg ccaaggagct ggagctgatg tttgggtgcc aggtggaagg
     1441 agatgcggct gagaccccac ccaggccaag gacccccggg aggcccctta gctcatacgg
     1501 aatggacagc cgacctccca tggcaatttt tgagttgttg gattacatag tcaacgagcc
     1561 tcctccaaaa ctgcccagtg gagtgttcag tctggaattt caagattttg tgaataaatg
     1621 cttaataaaa aaccccgcag agagagcaga tttgaagcaa ctcatggttc atgcttttat
     1681 caagagatct gatgctgagg aagtggattt tgcaggttgg ctctgctcca ccatcggcct
     1741 taaccagccc agcaccccaa cccatgctgc tggcgtccgg gcccgggatc caccggtcgc
     1801 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1861 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1921 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1981 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2041 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2101 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2161 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2221 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2281 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2341 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2401 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2461 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2521 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     2581 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2641 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2761 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2821 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2881 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2941 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3001 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3061 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3121 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3181 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3241 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3301 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3361 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3421 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3541 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3601 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3661 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3721 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3781 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3841 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3901 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3961 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4021 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4081 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4141 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4201 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4261 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4321 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4381 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4441 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4501 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4561 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4621 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4681 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4741 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4801 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4861 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4921 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4981 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5041 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5161 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5221 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5281 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5341 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5401 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5461 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5521 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5581 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5641 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5701 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5761 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5821 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     5881 tacggggtca ttagttcata gcccatatat ggagttcc
//
