LOCUS       sequence-128183-        9406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tandem tagged MAP3K15 in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9406)
  AUTHORS   Liebler
  TITLE     ASK_PLASMIDS
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9406)
  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: 
            "Unpublished"
COMMENT     Sequence Label: MAP3K15-TAP
FEATURES             Location/Qualifiers
     source          1..9406
                     /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"
     CDS             4903..4929
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4930..4959
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5000..5017)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5006..5230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5276..5704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5363..5382)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5573..5594
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5713..5733)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5718..6047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5898..6033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6114..6908
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6168..6187)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6778..6797
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7082..7203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7119..7138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7173..7192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7252..7268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7252..7268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7265..7287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7276..7292
                     /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(7300..7306,7307..7324,7325..7330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7345..7366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7483..7500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7654..8239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7734..7753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8410..9201,9202..9270))
                     /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     9033..9052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9271..9375)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ccgagctcgg atccactagt ccagtgtggt ggaattcatg
      961 gagagcggcg gtgggaatgc tccggccggg gccctcgggg cggcgagcga gtcccctcag
     1021 tgcccgccgc cgccgggggt ggagggcgcg gccgggccgg cggagcccga cggggcggcg
     1081 gagggcgcgg caggcggcag cggcgagggc gagagtgggg gcgggccgcg gcgggctctg
     1141 cgggcagtat acgtgcgcag tgagagctcc cagggcggcg cggccggcgg cccggaggct
     1201 ggggcgcggc agtgcctgct gcgggcctgc gaggccgagg gcgctcacct cacctccgtg
     1261 cccttcgggg agctggactt cggggagacg gccgtgctcg acgccttcta cgacgcagat
     1321 gttgctgtgg tagacatgag cgatgtctcc agacagcctt ccctcttcta ccatcttgga
     1381 gtccgagaaa gctttgacat ggccaataat gtgatcttgt accatgacac cgatgccgac
     1441 actgctctct ctttgaagga catggtaact caaaaaaaca cagcatccag tggaaattat
     1501 tatttcatcc catacatcgt gacaccgtgc gctgattatt tttgctgcga gagtgatgcc
     1561 cagagacgag cctccgagta catgcagccc aactgggaca acatcctggg cccgctgtgc
     1621 atgcctttgg tggacaggtt cattagcctc cttaaggaca tccacgtgac ctcatgtgtt
     1681 tattacaaag aaaccttgtt aaatgacatc cggaaagcca gagagaaata ccaaggtgag
     1741 gaactggcga aggagctagc tcggatcaag ctccgcatgg ataatactga ggttctgacc
     1801 tcagacatca tcattaactt actcctgtcc taccgtgata tccaggacta tgatgcgatg
     1861 gtgaagctgg tggaaacact ggagatgctg cctacgtgtg atttggccga tcagcataac
     1921 attaaattcc actatgcgtt tgcactgaat aggagaaaca gcacaggtga ccgtgagaag
     1981 gctctgcaga tcatgctcca ggttctgcag agctgtgatc acccgggccc cgacatgttc
     2041 tgcctgtgtg ggaggatcta caaggacatc ttcttggatt cagactgcaa agatgacacc
     2101 agccgcgaca gcgccattga gtggtatcgc aaagggtttg aactccagtc atccctctat
     2161 tcgggaatta atcttgcagt tttgctgatt gttgctggac aacaatttga aacttccttg
     2221 gaactaagga aaataggtgt ccggctgaac agtttgttgg gaagaaaagg gagcttggag
     2281 aaaatgaaca attactggga tgtgggtcag ttcttcagcg tcagcatgct ggcccatgat
     2341 gtcgggaaag ccgtccaggc agcagagagg ttgttcaaac tgaaacctcc agtctggtac
     2401 ctgcgatcat tagttcagaa cttgttacta attcggcgct tcaagaaaac cattattgaa
     2461 cactcgccca ggcaagagcg gctgaacttc tggttagata taatttttga ggcaacaaat
     2521 gaagtcacta atggactcag atttccagtt ctggtcatag agccaaccaa agtgtaccag
     2581 ccttcttatg tttccataaa caatgaagcc gaggagagaa cagtttcttt atggcatgtc
     2641 tcacccacag aaatgaaaca gatgcacgaa tggaatttta cagcctcttc cataaaggga
     2701 ataagcctat caaagtttga tgaaaggtgt tgttttcttt atgtccatga taattctgat
     2761 gactttcaaa tctacttttc caccgaagag cagtgcagta gatttttctc tttggtcaaa
     2821 gagatgataa ccaatacagc aggcagtacg gtggagctgg agggagagac cgatggagac
     2881 accttggagt atgagtatga ccatgatgca aatggtgaga gagttgtctt ggggaaaggc
     2941 acgtatggga ttgtgtatgc tggccgagat ctgagcaatc aagtgcgaat agccatcaaa
     3001 gaaatcccgg agagagatag caggtattct cagcctctgc acgaggagat agccctgcac
     3061 aagtacctta agcaccgcaa tatcgttcag tacctgggct ctgtttcaga gaacggctac
     3121 attaagatat ttatggagca ggtgcctgga ggaagccttt ctgctcttct gcgatccaaa
     3181 tgggggccga tgaaggaacc gacaatcaag ttttacacca aacagatcct ggagggcctt
     3241 aagtatcttc atgaaaacca gatcgtgcac agagacataa agggcgataa tgttctggtg
     3301 aacacctaca gcggagtggt gaaaatctcc gattttggaa cctcgaaacg tcttgcgggt
     3361 gtgaacccct gcacagagac ttttactggc accctgcagt acatggcacc tgagataatt
     3421 gaccaagggc ctcgcggata tggtgcccca gccgatatct ggtccctggg ctgcaccatc
     3481 attgagatgg ccaccagcaa gcctccgttc catgagcttg gtgagccgca ggcagccatg
     3541 ttcaaagtgg gcatgtttaa gatccaccct gagattccag aagccctttc agctgaagcc
     3601 cgagccttca ttttatcctg tttcgagcct gacccccaca aacgtgccac cactgctgag
     3661 ctactgagag agggtttctt aaggcaggtg aacaagggca agaagaaccg aattgccttc
     3721 aagccctcag aaggtccccg cggtgtcgtc ctggccctgc ccacacaggg agagcccatg
     3781 gccaccagca gcagcgagca cggctctgtc tccccagact ccgacgccca gcctgacgca
     3841 ctctttgaga ggacccgggc gcccaggcac caccttggcc acctcctcag tgttccagac
     3901 gagagctcag ccttggaaga ccggggcttg gcctcgtccc cggaggacag ggaccagggc
     3961 ctcttcctgc tacgcaagga cagtgagcgc cgtgccatcc tgtacaaaat cctctgggag
     4021 gagcagaacc aggtggcttc caacctgcag gagtgtgtgg cccagagttc cgaagagttg
     4081 catctctcag ttggacacat caagcaaatc attgggatcc tgagggactt catccgctcc
     4141 ccagagcacc gggtgatggc gaccacaata tcaaagctca aggtggacct ggactttgac
     4201 agctcgtcca tcagtcagat tcacctggtg ctgttcggat ttcaggatgc cgtaaataaa
     4261 attttgagga accacttaat taggccccac tggatgttcg cgatggacaa catcatccgc
     4321 cgagcggtgc aggccgcggt caccattctc atcccagagc tccgagccca ctttgagcct
     4381 acctgtgaga ctgaaggggt agataaggac atggatgaag cggaagaggg ctatccccca
     4441 gccaccggac ctggccagga ggcccagccc caccagcagc acctgagcct ccagctgggt
     4501 gagctcagac aggagaccaa cagacttttg gaacacctag ttgaaaaaga gagagagtac
     4561 cagaatcttc tgcggcaaac tctagaacag aaaactcaag aattgtatca ccttcagtta
     4621 aaattaaaat cgaattgtat tacagagaac ccagcaggcc cctacgggca gagaacagat
     4681 aaagagctta tagactggtt gcggctgcaa ggagctgatg caaagacaat tgaaaagatt
     4741 gttgaagagg gttatacact ttcggatatt cttaatgaga tcactaagga agatctaaga
     4801 taccttcgac tacggggtgg tctcctctgc agactctgga gtgcggtctc ccagtacaga
     4861 agggctcagg aggcctcaga aaccaaagac aaggctgtcg actatccgta tgatgttcct
     4921 gattatgctg aacagaaact gattagcgaa gaagatctgt agctcgagtc tagagggccc
     4981 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5041 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5101 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5161 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     5221 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     5281 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5341 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5401 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5461 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5521 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5581 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5641 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5701 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5821 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5881 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5941 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6001 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6061 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6121 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6181 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6241 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6301 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6361 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6421 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6481 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6541 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6601 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6661 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6721 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6781 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6841 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6901 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6961 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7021 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7081 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7201 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7261 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7321 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8161 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     8221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8341 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8401 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8461 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8521 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8581 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8641 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8701 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8761 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8821 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8881 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8941 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9001 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9061 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9121 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9181 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9241 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9301 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9361 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
