LOCUS       sequence-418254-        9388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of anti-Kv8.2 K+ channel (Rat) IgG2a 
            R-mAb. Derived from hybridoma N448/50..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9388)
  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 9388)
  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-Kv8.2 K+ channel [N448/50R]
FEATURES             Location/Qualifiers
     source          1..9388
                     /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        2390..2693
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2694..2897
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4139..4834
                     /codon_start=1
                     /product="CH2 and CH3 domains of the mouse IgG2a heavy 
                     chain plus the hinge region"
                     /label=Fc tag
                     /translation="PRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVT
                     CVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEF
                     KCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIY
                     VEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHT
                     TKSFSRTPGK"
     polyA_signal    4885..4940
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4939..4958)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7176..7194)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7262..7366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7367..7435,7436..8227)
                     /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(7585..7604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8398..8986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8887..8906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9140..9157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9274..9295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9310..9315,9316..9333,9334..9340)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9348..9364
                     /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     9353..9375
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9372..9388
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9372..9388
                     /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 gcagatgttt 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 atcagcagag tggaggctga ggatttggga gtttattatt
     1741 gctggcaagg tacacatttt cccacgttcg gtgctgggac caagctggag ctgaaacggg
     1801 ctgatgctgc accaactgta tccatcttcc caccatccag tgagcagtta acatctggag
     1861 gtgcctcagt cgtgtgcttc ttgaacaact tctaccccaa agacatcaat gtcaagtgga
     1921 agattgatgg cagtgaacga caaaatggcg tcctgaacag ttggactgat caggacagca
     1981 aagacagcac ctacagcatg agcagcaccc tcacgttgac caaggacgag tatgaacgac
     2041 ataacagcta tacctgtgag gccactcaca agacatcaac ttcacccatt gtcaagagct
     2101 tcaacaggaa tgagtgttag agacaaaggt cctgagacgc caccaccagc tccccagctc
     2161 catcctatct tcccttctaa ggtcttggag gcttccccac gagcgaccta ccactgttgc
     2221 ggtgctccaa acctcctccc cacctccttc tcctcctcct ccctttcctt ggcttttatc
     2281 atgctaatat ttgcagaaaa tattcaataa agtgagtctt tgcacttgat agttattaat
     2341 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2401 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2461 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2521 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
     2581 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
     2641 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2701 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2761 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     2821 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
     2881 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
     2941 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
     3001 actctcggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
     3061 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
     3121 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
     3181 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
     3241 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     3301 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctagcc
     3361 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     3421 ccactcccag gccaccatga agctgcctgt tctgctagtg gtgctgctat tgttcacgag
     3481 tccagcctca agcagtgaag tgatgctggt ggagtctggg ggaggcttag tgaagcctgg
     3541 agggtccctg aaactctcct gtgcagcctc tggattcact ttcagtaact ttgccctgtc
     3601 ttgggttcgt cagactccgg agaagaggct ggagtgggtc gcaaccatta gtagtggtgg
     3661 tagttacacc tactatccag acagtgtgaa ggggcgattc accatctcca gagacaatgc
     3721 caagaacacc ctgtatttgc aaatgagcag tctgaggtct gaggacacgg ccatgtattt
     3781 ctgtgcaagc gatggttact acgggtctta ctggggccaa gggactctgg tcactgtctc
     3841 tgcagcgcgc ccaacagccc catcggtcta tccactggcc cctgtgtgtg gagatacaac
     3901 tggctcctcg gtgactctag gatgcctggt caagggttat ttccctgagc cagtgacctt
     3961 gacctggaac tctggatccc tgtccagtgg tgtgcacacc ttcccagctg tcctgcagtc
     4021 tgacctctac accctcagca gctcagtgac tgtaacctcg agcacctggc ccagccagtc
     4081 catcacctgc aatgtggccc acccggcaag cagcaccaag gtggacaaga aaattgagcc
     4141 cagagggccc acaatcaagc cctgtcctcc atgcaaatgc ccagcaccta acctcttggg
     4201 tggaccatcc gtcttcatct tccctccaaa gatcaaggat gtactcatga tctccctgag
     4261 ccccatagtc acatgtgtgg tggtggatgt gagcgaggat gacccagatg tccagatcag
     4321 ctggtttgtg aacaacgtgg aagtacacac agctcagaca caaacccata gagaggatta
     4381 caacagtact ctccgggtgg tcagtgccct ccccatccag caccaggact ggatgagtgg
     4441 caaggagttc aaatgcaagg tcaacaacaa agacctccca gcgcccatcg agagaaccat
     4501 ctcaaaaccc aaagggtcag taagagctcc acaggtatat gtcttgcctc caccagaaga
     4561 agagatgact aagaaacagg tcactctgac ctgcatggtc acagacttca tgcctgaaga
     4621 catttacgtg gagtggacca acaacgggaa aacagagcta aactacaaga acactgaacc
     4681 agtcctggac tctgatggtt cttacttcat gtacagcaag ctgagagtgg aaaagaagaa
     4741 ctgggtggaa agaaatagct actcctgttc agtggtccac gagggtctgc acaatcacca
     4801 cacgactaag agcttctccc ggactccggg taaatgatct agacacgtga ttaattaagg
     4861 atcccccgac ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     4921 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga
     4981 tccctcggag atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta
     5041 cgacatctct gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca
     5101 acttgccaac tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt
     5161 ctctgactgt agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct
     5221 ttcatcctgg agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta
     5281 actcttggct gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga
     5341 agactacggg aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg
     5401 accctcaaga gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt
     5461 agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata
     5521 tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac
     5581 agcgatatct cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac
     5641 gagggtagtg aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg
     5701 aactctcctg aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg
     5761 ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc
     5821 cccagaataa aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa
     5881 ccctcacaaa ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt
     5941 taacaataga aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg
     6001 aatttatggc tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat
     6061 gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg
     6121 ggaaagagag tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa
     6181 attaaacggg gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt
     6241 ttgaaattgt ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact
     6301 gtaaaataag ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc
     6361 acttgcccac aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg
     6421 ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag
     6481 cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta
     6541 atgttgccat gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
     6601 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     6661 atatctgggt agtatatgct atcctaattt atatctgggt agcataggct atcctaatct
     6721 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct
     6781 gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct atcctaattt
     6841 atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     6901 ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     6961 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     7021 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     7081 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag
     7141 ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     7201 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7261 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7321 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7381 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7441 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7501 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7561 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     7621 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7681 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7741 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7801 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7861 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     7921 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7981 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8041 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8101 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8161 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8221 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8281 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8341 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8401 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8461 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8521 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8581 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8641 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     8701 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     8761 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8821 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8881 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8941 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9001 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     9061 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     9121 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     9181 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     9241 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     9301 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     9361 acaatttcac acaggaaaca gctatgac
//
