LOCUS       sequence-418273-        9415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of anti-MAP3K12 (Rat) subclass-switched
            IgG2b R-mAb. Derived from hybridoma N377/20..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9415)
  AUTHORS   Mitchell KG, Gong B, Hunter SS, Burkart-Waco D, Gavira-O'Neill CE, 
            Templeton KM, Goethel ME, Bzymek M, MacNiven LM, Murray KD, Settles 
            ML, Froenicke L, Trimmer JS
  TITLE     High-volume hybridoma sequencing on the NeuroMabSeq platform enables
            efficient generation of recombinant monoclonal antibodies and scFvs 
            for neuroscience research.
  JOURNAL   Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4.
  PUBMED    37758930
REFERENCE   2  (bases 1 to 9415)
  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: "Sci Rep."; 
            date: "2023-09-27"; volume: "13(1):16200. doi"; pages: " 
            10.1038/s41598-023-43233-4"
COMMENT     Sequence Label: Anti-MAP3K12 [N377/20R-2b]
FEATURES             Location/Qualifiers
     source          1..9415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        271..650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        651..854
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1398..1407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        2399..2702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2703..2906
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     polyA_signal    4912..4967
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4966..4985)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7203..7221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7289..7393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7394..7462,7463..8254)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7612..7631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8425..9013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8914..8933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9167..9184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9301..9322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9337..9342,9343..9360,9361..9367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9375..9391
                     /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     9380..9402
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9399..9415
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9399..9415
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
       61 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
      121 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
      181 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
      241 ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt
      301 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      361 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      421 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      481 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
      541 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
      601 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      661 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      721 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      781 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      841 atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc atcgctgtct
      901 gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct ttccagtact
      961 cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga cctgagcgag
     1021 tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc acagtcgcaa
     1081 ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt tctggcggag
     1141 gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt cgaggtgagg
     1201 tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag cgggcattac
     1261 ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccga
     1321 tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca caggtgtcca
     1381 ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg tccctgctgc
     1441 tctgcatgtc tggtgcggcc gcagatgttg tgatgaccca gactccactc actttgtcgg
     1501 ttaccattgg acaaccagcc tccatctctt gcaagtcaag tcagagcctc ttagatagtg
     1561 atggaaagac atatttgaat tggttgttac agaggccagg ccagtctcca aagcgcctaa
     1621 tctatctggt gtctaaactg gactctggag tccctgacag gttcactggc agtggatcag
     1681 ggacagattt cacactgaaa atcaggagag tggaggctga ggatttggga gtttattatt
     1741 gctggcaagg tacacatttt cctcagacgt tcggtggagg caccaagctg gaaatcaaac
     1801 gggctcgggc tgatgctgca ccaactgtat ccatcttccc accatccagt gagcagttaa
     1861 catctggagg tgcctcagtc gtgtgcttct tgaacaactt ctaccccaaa gacatcaatg
     1921 tcaagtggaa gattgatggc agtgaacgac aaaatggcgt cctgaacagt tggactgatc
     1981 aggacagcaa agacagcacc tacagcatga gcagcaccct cacgttgacc aaggacgagt
     2041 atgaacgaca taacagctat acctgtgagg ccactcacaa gacatcaact tcacccattg
     2101 tcaagagctt caacaggaat gagtgttaga gacaaaggtc ctgagacgcc accaccagct
     2161 ccccagctcc atcctatctt cccttctaag gtcttggagg cttccccacg agcgacctac
     2221 cactgttgcg gtgctccaaa cctcctcccc acctccttct cctcctcctc cctttccttg
     2281 gcttttatca tgctaatatt tgcagaaaat attcaataaa gtgagtcttt gcacttgata
     2341 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2401 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2461 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2521 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2581 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2641 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2701 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
     2761 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2821 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
     2881 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcctc actctcttcc
     2941 gcatcgctgt ctgcgagggc cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt
     3001 ctttccagta ctctcggatc ggaaacccgt cggcctccga acggtactcc gccaccgagg
     3061 gacctgagcg agtccgcatc gaccggatcg gaaaacctct cgagaaaggc gtctaaccag
     3121 tcacagtcgc aaggtaggct gagcaccgtg gcgggcggca gcgggtggcg gtcggggttg
     3181 tttctggcgg aggtgctgct gatgatgtaa ttaaagtagg cggtcttgag acggcggatg
     3241 gtcgaggtga ggtgtggcag gcttgagatc cagctgttgg ggtgagtact ccctctcaaa
     3301 agcgggcatt acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt
     3361 cacctagccc gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc
     3421 cacaggtgtc cactcccagg ccaccatgaa gctgcctgtt ctgctagtgg tgctgctatt
     3481 gttcacgagt ccagcctcaa gcagtcagat ccagttggtg cagtctggac ctgagctgaa
     3541 gaagcctgga gagactgtca agctctcctg caaggcttct ggatacacct tcacaaactt
     3601 tggaatgaac tgggtgaagc aggatccagg aaagggttta aagtggatgg gctggataaa
     3661 caccttcact ggagagccaa cctatgctga tgacttcaag ggacggtttg ccttctcttt
     3721 ggaaacctct gccaacactg cctatttgca gatcatcaac ctcaaaagtg aggacatggc
     3781 tacatatttc tgtgccatct ttaggtacga cggtccttac tggggccaag ggactctggt
     3841 cactgtctct acagcgcgcc caacaccccc ctcagtgtac ccccttgctc cagggtgcgg
     3901 cgacactact ggatctagcg ttacactggg ctgtttggtg aaagggtatt ttccagagtc
     3961 agttactgta acctggaaca gcggaagtct ttcttcaagt gtgcatacat tccctgcttt
     4021 gcttcagtcc ggcctgtaca ccatgtcttc ttctgtgact gtgcctagtt ccacctggcc
     4081 atcccagact gtcacatgct cagtggccca tccagccagc agcacaaccg tggacaagaa
     4141 actagagcca tcaggtccca tcagcacgat aaacccatgc ccaccctgca aggaatgtca
     4201 caaatgccct gcaccaaacc tggaaggggg tccttccgtg ttcatatttc caccgaatat
     4261 taaagatgtc ctcatgatta gcctcacccc caaagtgaca tgtgtagttg tagatgtttc
     4321 ggaggatgat cctgatgtcc agatctcctg gtttgtaaac aatgtcgaag tccacaccgc
     4381 ccagacgcaa acacacagag aagactacaa cagcaccatc cgagtggtca gcacactccc
     4441 gattcagcac caggactgga tgagcggtaa ggagttcaag tgtaaagtca acaacaagga
     4501 tctgcctagt cctattgaaa gaaccatcag taagatcaaa ggactggtga gggccccgca
     4561 agtttatatc ctgcctcctc ccgccgagca gttatctcgc aaggacgttt ccctcacttg
     4621 tctggtagta ggctttaacc ctggcgacat ctccgtggag tggacgagca atgggcacac
     4681 agaggagaac tacaaggaca ctgcgcccgt gctagattca gacggctctt acttcatata
     4741 tagcaaactg aatatgaaaa ccagcaagtg ggagaagaca gactctttct cctgcaatgt
     4801 gaggcatgaa ggattgaaga attattacct gaagaagacc atttcacgga gtcccggaaa
     4861 atgatctaga cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga
     4921 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4981 atgggagggc aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc
     5041 ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc
     5101 atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca
     5161 cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat
     5221 gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac
     5281 tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg
     5341 ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga
     5401 ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata
     5461 aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat
     5521 ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt
     5581 agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt
     5641 tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag
     5701 tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc
     5761 tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg
     5821 ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg
     5881 tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt
     5941 gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc
     6001 gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt
     6061 gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca
     6121 tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc
     6181 cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata
     6241 aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc
     6301 ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt
     6361 aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg
     6421 gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag
     6481 gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca
     6541 cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata
     6601 tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata
     6661 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata
     6721 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     6781 tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga
     6841 ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg
     6901 gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     6961 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     7021 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     7081 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     7141 ggtagcatat gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa
     7201 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     7261 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7621 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     7681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7921 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9181 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     9241 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     9301 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     9361 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgac
//
