LOCUS       sequence-418366-        6910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of Botch scFv. Derived from hybridoma 
            N116/14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  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 6910)
  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: Botch scFv [N116/14]
FEATURES             Location/Qualifiers
     source          1..6910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(49..840,841..909))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     672..691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(910..1014)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1089..1108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1280..1659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1660..1863
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1814..1834
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1860..1884
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1983..1992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2793..2819
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2844..2861
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2917..3505
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2970..2990)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3388..3399)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3526..3545)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3570..3618
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3820..4248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4117..4138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4257..4277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4262..4591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4442..4577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4504..4523
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4658..5452
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4712..4731)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5322..5341
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5628..5749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5665..5684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5719..5738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5798..5814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5798..5814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5811..5833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5822..5838
                     /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)."
     promoter        complement(join(5846..5852,5853..5870,5871..5876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5891..5912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6029..6046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6200..6788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6280..6299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
       61 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      121 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      181 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      241 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      301 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      361 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      421 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      481 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      541 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      601 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      661 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      721 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      781 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      841 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      901 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      961 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1021 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1081 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     1141 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     1201 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     1261 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     1321 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     1381 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     1441 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1501 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     1561 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     1621 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     1681 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     1741 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     1801 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     1861 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     1921 agaagacacc gggaccgatc cagcctccgg actctagagg atcgaaccct tgaattcccg
     1981 ccgccaccat gggctggagc tgtattattc tcttcttggt ggcgaccgca actggagtac
     2041 attcccaagt gcaactccaa caaagcggcg ccgaactcat gaaacccgga gcgtctgtca
     2101 aaattagttg taaagccacg ggttatactt ttaacaatta ttggattgaa tggatcaaac
     2161 aacgtcccgg ccacggactc gaatggatcg gcgaaatctt gccggggtca gactctaatt
     2221 ataacgaaaa gtttaaagga aaagcaacgt ttacgactga cacgagcagc aatactgcat
     2281 atatccagct gtcatccctc acttcagaag atagtgctgt atactattgc tcccggggtt
     2341 acggctattt tacatggttt gcctattggg gacagggaac actcgtaaca gtgagtgctg
     2401 gtggtggtgg atcaggtggc ggcggttcag gtggtggtgg gtcaggcggt ggatccgacg
     2461 tggtcatgac acagacccct ttaagcctgc cagtttccct gggcgaccag gcaagcatta
     2521 gttgtcgtag ctcccaaagt ctggtgcata gcaacggtaa tacatacttg cactggtatc
     2581 tccagaaacc tggtcaatca cctaaactgc tcatttataa ggtaagcaat cgattcagcg
     2641 gagttcccga tcgcttttcc ggttcaggtt ccggcacgga ctttactctt aaaatttctc
     2701 gtgtagaagc agaagacctt gggatctact tttgttccca ggggacccac gtcccatata
     2761 cctttggtgg cggaacaaaa cttgagatta agtatcccta tgatgtacct gactatgctt
     2821 ccggcctgcc cgaaacaggc ggtcatcatc atcatcatca ctaataagct taagggttcg
     2881 atccctaccg gttagtaatg agtttgatat ctcgacaatc aacctctgga ttacaaaatt
     2941 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3001 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3061 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3121 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3181 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3241 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3301 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     3361 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3421 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3481 atctcccttt gggccgcctc cccgcctgga acgggggagg ctaactgaaa cacggaagga
     3541 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3601 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     3661 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     3721 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     3781 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4021 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4141 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4201 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4261 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4321 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4441 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4501 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4561 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4621 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4681 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4741 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4801 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4861 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4921 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4981 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5041 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5101 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5161 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5221 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5281 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5341 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5401 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5461 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5521 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5581 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5641 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5701 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5761 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5821 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5881 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5941 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6001 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6061 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6901 attaaaaatg
//
