LOCUS       sequence-418249-        9385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of anti-Kv6.1 K+ channel (Rat) IgG2a 
            R-mAb. Derived from hybridoma N474/27..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9385)
  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 9385)
  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-Kv6.1 K+ channel [N474/27R]
FEATURES             Location/Qualifiers
     source          1..9385
                     /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        2393..2696
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2697..2900
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4136..4831
                     /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    4882..4937
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4936..4955)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7173..7191)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7259..7363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7364..7432,7433..8224)
                     /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(7582..7601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8395..8983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8884..8903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9137..9154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9271..9292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9307..9312,9313..9330,9331..9337)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9345..9361
                     /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     9350..9372
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9369..9385
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9369..9385
                     /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 atcagcagag tggaggctga ggatttggga gtttattttt
     1741 gctggcaagg tacacatttt cctcacacgt tcgggggggg gaccaagctg gaaataaaac
     1801 gggctgatgc tgcaccaact gtatccatct tcccaccatc cagtgagcag ttaacatctg
     1861 gaggtgcctc agtcgtgtgc ttcttgaaca acttctaccc caaagacatc aatgtcaagt
     1921 ggaagattga tggcagtgaa cgacaaaatg gcgtcctgaa cagttggact gatcaggaca
     1981 gcaaagacag cacctacagc atgagcagca ccctcacgtt gaccaaggac gagtatgaac
     2041 gacataacag ctatacctgt gaggccactc acaagacatc aacttcaccc attgtcaaga
     2101 gcttcaacag gaatgagtgt tagagacaaa ggtcctgaga cgccaccacc agctccccag
     2161 ctccatccta tcttcccttc taaggtcttg gaggcttccc cacgagcgac ctaccactgt
     2221 tgcggtgctc caaacctcct ccccacctcc ttctcctcct cctccctttc cttggctttt
     2281 atcatgctaa tatttgcaga aaatattcaa taaagtgagt ctttgcactt gatagttatt
     2341 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     2401 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2461 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2521 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc
     2581 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2641 tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     2701 tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     2761 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     2821 caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg
     2881 aggtctatat aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg
     2941 ctgtctgcga gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc
     3001 agtactctcg gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg
     3061 agcgagtccg catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag
     3121 tcgcaaggta ggctgagcac cgtggcgggc ggcagcgggt ggcggtcggg gttgtttctg
     3181 gcggaggtgc tgctgatgat gtaattaaag taggcggtct tgagacggcg gatggtcgag
     3241 gtgaggtgtg gcaggcttga gatccagctg ttggggtgag tactccctct caaaagcggg
     3301 cattacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcaccta
     3361 gcccgatctg gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg
     3421 tgtccactcc caggccacca tgaagctgcc tgttctgcta gtggtgctgc tattgttcac
     3481 gagtccagcc tcaagcagtg aggttcagtt ccagcagtct gggactgtgc tggcaaggcc
     3541 tgggacttca gtgaagatgt cctgcaaggc ttctggctac acctttacca gctactggat
     3601 gcactgggta aaacagaggc ctggacaggg tctagaatgg attggcgcta tttatcctgg
     3661 aaatagtgat gctgactaca accagaagtt caagggcaag gccaaactga ctgctgtcac
     3721 atccaccagc actgcctaca tggagctcag cagcctgaca aatgaagact ctgcggtcta
     3781 ttactgtaca agaaatagag gaggctactg gggccaaggc accactctca cagtctcctc
     3841 agcgcgccca acagccccat cggtctatcc actggcccct gtgtgtggag atacaactgg
     3901 ctcctcggtg actctaggat gcctggtcaa gggttatttc cctgagccag tgaccttgac
     3961 ctggaactct ggatccctgt ccagtggtgt gcacaccttc ccagctgtcc tgcagtctga
     4021 cctctacacc ctcagcagct cagtgactgt aacctcgagc acctggccca gccagtccat
     4081 cacctgcaat gtggcccacc cggcaagcag caccaaggtg gacaagaaaa ttgagcccag
     4141 agggcccaca atcaagccct gtcctccatg caaatgccca gcacctaacc tcttgggtgg
     4201 accatccgtc ttcatcttcc ctccaaagat caaggatgta ctcatgatct ccctgagccc
     4261 catagtcaca tgtgtggtgg tggatgtgag cgaggatgac ccagatgtcc agatcagctg
     4321 gtttgtgaac aacgtggaag tacacacagc tcagacacaa acccatagag aggattacaa
     4381 cagtactctc cgggtggtca gtgccctccc catccagcac caggactgga tgagtggcaa
     4441 ggagttcaaa tgcaaggtca acaacaaaga cctcccagcg cccatcgaga gaaccatctc
     4501 aaaacccaaa gggtcagtaa gagctccaca ggtatatgtc ttgcctccac cagaagaaga
     4561 gatgactaag aaacaggtca ctctgacctg catggtcaca gacttcatgc ctgaagacat
     4621 ttacgtggag tggaccaaca acgggaaaac agagctaaac tacaagaaca ctgaaccagt
     4681 cctggactct gatggttctt acttcatgta cagcaagctg agagtggaaa agaagaactg
     4741 ggtggaaaga aatagctact cctgttcagt ggtccacgag ggtctgcaca atcaccacac
     4801 gactaagagc ttctcccgga ctccgggtaa atgatctaga cacgtgatta attaaggatc
     4861 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     4921 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc
     4981 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     5041 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     5101 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     5161 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     5221 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     5281 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     5341 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     5401 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     5461 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     5521 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     5581 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     5641 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     5701 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     5761 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     5821 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     5881 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     5941 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     6001 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     6061 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     6121 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     6181 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     6241 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     6301 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     6361 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
     6421 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
     6481 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
     6541 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     6601 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     6661 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     6721 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     6781 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata
     6841 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     6901 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     6961 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     7021 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     7081 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg
     7141 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     7201 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     7621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     7921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     9061 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     9121 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     9181 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     9241 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     9301 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     9361 atttcacaca ggaaacagct atgac
//
