LOCUS       sequence-431009-        8743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus vector for expression of MEK1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8743)
  TITLE     Der Lab MEK1 Plasmids 2024
REFERENCE   2  (bases 1 to 8743)
  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"
COMMENT     Sequence Label: WT-MEK1-V5
FEATURES             Location/Qualifiers
     source          1..8743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..226
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        38..234
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      85..220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             266..868
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(266..285)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     762..782
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    911..1028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1086..1586
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1505..1523
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2780..2821
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2846..3434
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2899..2919)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3317..3328)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3506..3739
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3817..3938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3854..3873)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3908..3927
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3978..4113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4133..4152)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4134..4152)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4162..4179)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4162..4178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4171..4193)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4320..4775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4617..4638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4801..4905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4906..4974,4975..5766)
                     /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(5124..5143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5937..6525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6426..6445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6679..6696
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6813..6834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6849..6854,6855..6872,6873..6879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6887..6903
                     /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)."
     primer_bind     6892..6914
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6911..6927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6911..6927
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6946..6966
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6948..6966
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6994..7220
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7221..7401
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7448..7573
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8066..8299
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8484..8528
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 tccccaggct ccccagcagg cagaagtatg caaagcatgc gtctcaatta gtcagcaacc
       61 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
      181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
      241 tctagaggta ccaccggagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
      301 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
      361 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
      421 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
      481 cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
      541 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
      601 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
      661 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
      721 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
      781 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
      841 catgacccgc aagcccggtg ccagttaacg aattctagat cttgagacaa atggcagtat
      901 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
      961 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     1021 aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggcct
     1081 gcaggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc tgctctgggc
     1141 gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc acgtccgttc
     1201 gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg cttcctgctc
     1261 cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga caaacggaag
     1321 ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc
     1381 gaccgcgatg ggctgtggcc aatagcggct gctcagcagg gcgcgccgag agcagcggcc
     1441 gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg
     1501 cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg
     1561 accgaatcac cgacctctct ccccaggggc tagcatgccc aagaagaagc cgacgcccat
     1621 ccagctgaac ccggcccccg acggctctgc agttaacggg accagctctg cggagaccaa
     1681 cttggaggcc ttgcagaaga agctggagga gctagagctt gatgagcagc agcgaaagcg
     1741 ccttgaggcc tttcttaccc agaagcagaa ggtgggagaa ctgaaggatg acgactttga
     1801 gaagatcagt gagctggggg ctggcaatgg cggtgtggtg ttcaaggtct cccacaagcc
     1861 ttctggcctg gtcatggcca gaaagctaat tcatctggag atcaaacccg caatccggaa
     1921 ccagatcata agggagctgc aggttctgca tgagtgcaac tctccgtaca tcgtgggctt
     1981 ctatggtgcg ttctacagcg atggcgagat cagtatctgc atggagcaca tggatggagg
     2041 ttctctggat caagtcctga agaaagctgg aagaattcct gaacaaattt taggaaaagt
     2101 tagcattgct gtaataaaag gcctgacata tctgagggag aagcacaaga tcatgcacag
     2161 agatgtcaag ccttccaaca tcctagtcaa ctcccgtggg gagatcaagc tctgtgactt
     2221 tggggtcagc gggcagctca tcgactccat ggccaactcc ttcgtgggca caaggtccta
     2281 catgtcgcca gaaagactcc aggggactca ttactctgtg cagtcagaca tctggagcat
     2341 gggactgtct ctggtagaga tggcggttgg gaggtatccc atccctcctc cagatgccaa
     2401 ggagctggag ctgatgtttg ggtgccaggt ggaaggagat gcggctgaga ccccacccag
     2461 gccaaggacc cccgggaggc cccttagctc atacggaatg gacagccgac ctcccatggc
     2521 aatttttgag ttgttggatt acatagtcaa cgagcctcct ccaaaactgc ccagtggagt
     2581 gttcagtctg gaatttcaag attttgtgaa taaatgctta ataaaaaacc ccgcagagag
     2641 agcagatttg aagcaactca tggttcatgc ttttatcaag agatctgatg ctgaggaagt
     2701 ggattttgca ggttggctct gctccaccat cggccttaac cagcccagca ccccaaccca
     2761 tgctgctggc gtcaccggtg gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2821 gtagtaatga ccgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     2881 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2941 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3001 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3061 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3121 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3181 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3241 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3301 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3361 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3421 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     3481 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     3541 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     3601 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     3661 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     3721 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     3781 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     3841 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3901 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     3961 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4021 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4081 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     4141 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4201 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4261 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4741 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4801 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4861 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4921 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4981 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5041 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5101 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5161 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5221 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5281 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5341 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5401 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5461 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5521 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5581 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5641 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5701 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5761 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5821 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5881 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5941 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6001 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6061 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6121 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6181 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6241 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6301 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6361 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6421 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6481 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6541 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6601 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6661 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6721 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6781 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6841 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     6901 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     6961 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     7021 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     7081 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     7141 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     7201 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     7261 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     7321 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     7381 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     7441 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     7501 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     7561 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     7621 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     7681 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     7741 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     7801 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     7861 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     7921 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     7981 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     8041 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     8101 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     8161 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     8221 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     8281 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     8341 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     8401 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     8461 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     8521 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     8581 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     8641 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     8701 ccatgcattg catctcaatt agtcagcaac caggtgtgga aag
//
