LOCUS       sequence-418369-        9361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of anti-VAChT (Rat) subclass-switched 
            IgG1 R-mAb. Derived from hybridoma N425/45..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9361)
  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 9361)
  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-VAChT [N425/45R-1]
FEATURES             Location/Qualifiers
     source          1..9361
                     /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        2378..2681
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2682..2885
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     polyA_signal    4858..4913
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4912..4931)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7149..7167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7235..7339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7340..7408,7409..8200)
                     /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(7558..7577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8371..8959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8860..8879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9113..9130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9247..9268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9283..9288,9289..9306,9307..9313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9321..9337
                     /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     9326..9348
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9345..9361
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9345..9361
                     /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 gcagacatcc agatgaacca gtctccatcc agtctgtctg
     1501 catcccttgg agacacaatt accatcactt gccatgccag tcagaacatt aatgtttggt
     1561 taagctggta ccagcagaaa ccaagaaata ttcctaaact gttgatctat aaggcttcca
     1621 acttgcacac aggcgtccca tcaaggttta gtggcagtgg atctggaaca gatttcacat
     1681 taaccatcag cagcctgcag cctgaagaca ttgccactta ctactgtcaa cagggtcaaa
     1741 gttatcctct cacgttcggt gctgggacca agctggagct gaaacgggct gatgctgcac
     1801 caactgtatc catcttccca ccatccagtg agcagttaac atctggaggt gcctcagtcg
     1861 tgtgcttctt gaacaacttc taccccaaag acatcaatgt caagtggaag attgatggca
     1921 gtgaacgaca aaatggcgtc ctgaacagtt ggactgatca ggacagcaaa gacagcacct
     1981 acagcatgag cagcaccctc acgttgacca aggacgagta tgaacgacat aacagctata
     2041 cctgtgaggc cactcacaag acatcaactt cacccattgt caagagcttc aacaggaatg
     2101 agtgttagag acaaaggtcc tgagacgcca ccaccagctc cccagctcca tcctatcttc
     2161 ccttctaagg tcttggaggc ttccccacga gcgacctacc actgttgcgg tgctccaaac
     2221 ctcctcccca cctccttctc ctcctcctcc ctttccttgg cttttatcat gctaatattt
     2281 gcagaaaata ttcaataaag tgagtctttg cacttgatag ttattaatag taatcaatta
     2341 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2401 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2461 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2521 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
     2581 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
     2641 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2701 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2761 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
     2821 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2881 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc
     2941 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tctcggatcg
     3001 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcga gtccgcatcg
     3061 accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca aggtaggctg
     3121 agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga ggtgctgctg
     3181 atgatgtaat taaagtaggc ggtcttgaga cggcggatgg tcgaggtgag gtgtggcagg
     3241 cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatta cttctgcgct
     3301 aagattgtca gtttccaaaa acgaggagga tttgatattc acctagcccg atctggccat
     3361 acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     3421 caccatgaag ctgcctgttc tgctagtggt gctgctattg ttcacgagtc cagcctcaag
     3481 cagtcagatc cagttggtgc agtctggacc tgagctgaag aagcctggag agacagtcaa
     3541 gatctcctgc aaggcttctg gatatagttt cacaaactat ggaatgaact gggtgaagca
     3601 ggctccagga aagggtttaa agtggatggc ctggataaac acctacactg gagagccaac
     3661 atatgctgat gacttcaagg gacggtttgc cttctctttg gaaacctctg ccagcactgc
     3721 ctatttgcag atcaacaacc tcaaaaaaga ggacacggct acatatttct gcgcaaacct
     3781 atatagggac tactttgact actggggcca aggcaccact ctcacagtct cctcagcgcg
     3841 cccgacaccc ccatctgtct atccactggc ccctggatct gctgcccaaa ctaactccat
     3901 ggtgaccctg ggatgcctgg tcaagggcta tttccctgag ccagtgacag tgacctggaa
     3961 ctctggatcc ctgtccagcg gtgtgcacac cttcccagct gtcctgcagt ctgacctcta
     4021 cactctgagc agctcagtga ctgtcccctc cagcacctgg cccagcgaga ccgtcacctg
     4081 caacgttgcc cacccggcca gcagcaccaa ggtggacaag aaaattgtgc ccagggattg
     4141 tggttgtaag ccttgcatat gtacagtccc agaagtatca tctgtcttca tcttcccccc
     4201 aaagcccaag gatgtgctca ccattactct gactcctaag gtcacgtgtg ttgtggtaga
     4261 catcagcaag gatgatcccg aggtccagtt cagctggttt gtagatgatg tggaggtgca
     4321 cacagctcag acgcaacccc gggaggagca gttcaacagc actttccgct cagtcagtga
     4381 acttcccatc atgcaccagg actggctcaa tggcaaggag ttcaaatgca gggtcaacag
     4441 tgcagctttc cctgccccca tcgagaaaac catctccaaa accaaaggca gaccgaaggc
     4501 tccacaggtg tacaccattc cacctcccaa ggagcagatg gccaaggata aagtcagtct
     4561 gacctgcatg ataacagact tcttccctga agacattact gtggagtggc agtggaatgg
     4621 gcagccagcg gagaactaca agaacactca gcccatcatg gacacagatg gctcttactt
     4681 cgtctacagc aagctcaatg tgcagaagag caactgggag gcaggaaata ctttcacctg
     4741 ctctgtgtta catgagggcc tgcacaacca ccatactgag aagagcctct cccactctcc
     4801 tggtaaatga tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat
     4861 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     4921 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat
     4981 cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg
     5041 cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat
     5101 gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa
     5161 atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct
     5221 gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat
     5281 gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca
     5341 atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg
     5401 tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata
     5461 tactatccag actaacccta attcaatagc atatgttacc caacgggaag catatgctat
     5521 cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt
     5581 cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa
     5641 gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct
     5701 tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt
     5761 gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt
     5821 cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat
     5881 actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga
     5941 caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc aacacataat
     6001 cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt
     6061 gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc
     6121 ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg
     6181 cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt
     6241 cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct
     6301 gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac
     6361 cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat
     6421 ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca
     6481 tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc
     6541 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
     6601 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     6661 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     6721 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa
     6781 tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa
     6841 tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     6901 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     6961 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     7021 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     7081 tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag
     7141 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     7201 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7321 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7441 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7501 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7561 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     7621 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7741 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7861 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7921 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     8401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     8521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     8761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     9001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     9061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     9121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     9181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     9241 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     9301 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     9361 c
//
