LOCUS       sequence-418257-        9400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of anti-Versican (Rat) IgG2a R-mAb. 
            Derived from hybridoma N351/24..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9400)
  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 9400)
  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-Versican [N351/24R]
FEATURES             Location/Qualifiers
     source          1..9400
                     /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             4151..4846
                     /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    4897..4952
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4951..4970)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7188..7206)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7274..7378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7379..7447,7448..8239)
                     /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(7597..7616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8410..8998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8899..8918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9152..9169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9286..9307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9322..9327,9328..9345,9346..9352)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9360..9376
                     /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     9365..9387
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9384..9400
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9384..9400
                     /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 gcagatattg tgatgaccca aactccactc tccctgtctg
     1501 tcagtcttgg agatcaagcc tccatctctt gcagatctag tcagagcctt ttacacagta
     1561 atggaaacac ctatttacat tggtacctgc agaagccagg ccagtctcca aagctcctga
     1621 tctacaaagt ttccaaccga ttttctgggg tcccagacag gttcagtggc agtggatcag
     1681 ggacagattt cacactcaag atcagcagag tggaggctga ggatctggga gtttatttct
     1741 gctctcaaac tacacatgtt ccattcacgt tcggctcggg gacaaagttg 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 aggtccagct gcagcagtct ggacctgaac tggtaaagcc
     3541 tggggcttca gtgaagatgt cctgcagggc ttctggatac acattcacta gctatgttat
     3601 tctctgggtg aagcagaagc ctgggcaggg ccttgagtgg attggattta ttcatcctta
     3661 caatgatggt actaagtaca atgagaagtt caagggcaag gccacactga cttcagacaa
     3721 atcctccagc acagcctaca tggagctcag cagcctgacc tctgaggact ctgcggtcta
     3781 ttactgtgca agatgggggg actacggggt agtagccgac ttctggggcc aaggcaccac
     3841 tctcacagtc tcctcagcgc gcccaacagc cccatcggtc tatccactgg cccctgtgtg
     3901 tggagataca actggctcct cggtgactct aggatgcctg gtcaagggtt atttccctga
     3961 gccagtgacc ttgacctgga actctggatc cctgtccagt ggtgtgcaca ccttcccagc
     4021 tgtcctgcag tctgacctct acaccctcag cagctcagtg actgtaacct cgagcacctg
     4081 gcccagccag tccatcacct gcaatgtggc ccacccggca agcagcacca aggtggacaa
     4141 gaaaattgag cccagagggc ccacaatcaa gccctgtcct ccatgcaaat gcccagcacc
     4201 taacctcttg ggtggaccat ccgtcttcat cttccctcca aagatcaagg atgtactcat
     4261 gatctccctg agccccatag tcacatgtgt ggtggtggat gtgagcgagg atgacccaga
     4321 tgtccagatc agctggtttg tgaacaacgt ggaagtacac acagctcaga cacaaaccca
     4381 tagagaggat tacaacagta ctctccgggt ggtcagtgcc ctccccatcc agcaccagga
     4441 ctggatgagt ggcaaggagt tcaaatgcaa ggtcaacaac aaagacctcc cagcgcccat
     4501 cgagagaacc atctcaaaac ccaaagggtc agtaagagct ccacaggtat atgtcttgcc
     4561 tccaccagaa gaagagatga ctaagaaaca ggtcactctg acctgcatgg tcacagactt
     4621 catgcctgaa gacatttacg tggagtggac caacaacggg aaaacagagc taaactacaa
     4681 gaacactgaa ccagtcctgg actctgatgg ttcttacttc atgtacagca agctgagagt
     4741 ggaaaagaag aactgggtgg aaagaaatag ctactcctgt tcagtggtcc acgagggtct
     4801 gcacaatcac cacacgacta agagcttctc ccggactccg ggtaaatgat ctagacacgt
     4861 gattaattaa ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg
     4921 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     4981 atttggtcga gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac
     5041 taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag
     5101 ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa
     5161 cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa
     5221 cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg
     5281 cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc
     5341 aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct
     5401 accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa
     5461 ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa
     5521 ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg
     5581 gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg
     5641 tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag
     5701 gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta
     5761 atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt
     5821 tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg
     5881 acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat
     5941 tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt
     6001 ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg
     6061 ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta
     6121 tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta
     6181 acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg
     6241 ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg
     6301 cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac
     6361 tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag
     6421 taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca
     6481 cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag
     6541 cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg
     6601 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     6661 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
     6721 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     6781 ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg
     6841 ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat
     6901 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag cataggctat
     6961 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     7021 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     7081 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     7141 cctcatgata agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg
     7201 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7261 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7321 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7381 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7441 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7501 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7561 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7621 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7681 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7741 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7801 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7861 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7921 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7981 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     8041 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     8101 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     8161 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8221 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8281 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8341 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8521 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8641 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8701 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8761 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8821 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8881 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8941 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     9001 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     9061 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     9121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     9181 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     9241 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     9301 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     9361 tgtgagcgga taacaatttc acacaggaaa cagctatgac
//
