LOCUS       sequence-420247-        6904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of TRIP8b (exon 4) scFv. Derived from 
            hybridoma N212/3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6904)
  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 6904)
  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: TRIP8b (exon 4) scFv [N212/3]
FEATURES             Location/Qualifiers
     source          1..6904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..36)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    61..109
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      311..739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(398..417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     608..629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(748..768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        753..1082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      933..1068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     995..1014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1149..1943
                     /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(1203..1222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1813..1832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2119..2240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2156..2175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2210..2229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2289..2305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2289..2305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2302..2324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2313..2329
                     /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(2337..2343,2344..2361,2362..2367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2382..2403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2520..2537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2691..3279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2771..2790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3450..4241,4242..4310))
                     /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     4073..4092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4311..4415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4490..4509)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4681..5060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5061..5264
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5215..5235
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5261..5285
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5384..5393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6191..6217
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    6312..6900
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6365..6385)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6783..6794)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
       61 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
      121 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
      181 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
      241 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
      301 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      661 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      721 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
      781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
      841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
      961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     1021 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     1081 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     1141 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     1201 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     1261 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     1321 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     1381 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     1441 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     1501 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     1561 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     1621 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     1681 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     1741 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     1801 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     1861 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     1921 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     1981 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     2041 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     2101 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2221 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     2281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2341 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2881 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3121 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3301 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3361 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3421 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3481 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3541 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3601 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3661 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3721 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3781 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3841 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3901 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3961 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4021 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4081 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4141 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4201 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4261 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4321 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4381 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4441 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     4501 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     4561 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     4621 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     4681 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     4741 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4801 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4861 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4921 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4981 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     5041 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     5101 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     5161 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     5221 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     5281 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     5341 ccagcctccg gactctagag gatcgaaccc ttgaattccc gccgccacca tgggctggag
     5401 ctgtattata ctctttcttg tagctaccgc gacaggagtt cattcccaag tacaattaca
     5461 acaatccggt gccgaattgg tcaaaccagg cgcaagcgtt aaaataagct gtcgcgcctc
     5521 aggttatacg tttagcactt attggattca atgggtcaaa caacggccag gccacgggtt
     5581 agaatggatc ggggaaattc tcccaggcaa catctcaaca aattataacg aaaagtttaa
     5641 aggaaaagca acttttacgg ccgacacttc atcaaatacc gcttatatgc agctgtccag
     5701 tctcccgagc gaagattcag ctgtgtacta ttgcgctcgt ctgcggttgg ggtacgcaat
     5761 ggattattgg ggccagggta cgagcgtgac ggtgagttcc ggcggaggag ggtcaggtgg
     5821 tggtggatca ggtggggggg ggtcaggtgg cgggagcgat attgtactca cacagagtcc
     5881 cgcatccctt gctgtcagtc tcggccaacg cgcaactatt agttgtaggg cttctgagtc
     5941 cgtggactct tacggtaaca acttcatgca ttggtatcaa caaaagcccg gccaaccgcc
     6001 taagctgctt atttaccggg cctcaaatct ggagagtggc attccggcta gattttccgg
     6061 gagcggaagc cgcactgatt ttacgttgac tataaacccg gttgaagccg acgacgtggc
     6121 gacttactat tgccaacaga acaacgaaga cccttggacc tttggcggcg gaacaaaatt
     6181 ggagattaag tatccatatg atgtccctga ctatgcttca ggcctgccag aaactggcgg
     6241 tcatcatcat catcactaat aagcttaagg gttcgatccc taccggttag taatgagttt
     6301 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6361 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6421 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6481 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6541 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6601 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6661 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     6721 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6781 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6841 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6901 ctgg
//
