LOCUS       sequence-418365-        6910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of Pan-QKI scFv. Derived from hybridoma
            N147/6..
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: Pan-QKI scFv [N147/6]
FEATURES             Location/Qualifiers
     source          1..6910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      789..1377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1278..1297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1531..1548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1665..1686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1701..1706,1707..1724,1725..1731)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1739..1755
                     /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     1744..1766
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1763..1779
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1763..1779
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1828..1949
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1839..1858)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1893..1912
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2125..2919)
                     /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(2236..2255)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2846..2865
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2986..3315)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3000..3135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3054..3073)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3300..3320
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3329..3757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3439..3460)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3651..3670
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3959..4007
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4032..4051
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     misc_feature    4072..4660
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4178..4189
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4587..4607
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4716..4733)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4758..4784)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      5585..5594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5693..5717)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(5714..5917)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5743..5763)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5918..6297
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6469..6488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6668..6736,6737..6910,1..618)
                     /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     complement(6886..6905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
       61 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      301 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      601 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      661 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      781 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      841 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      901 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      961 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1021 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1081 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1141 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1201 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1261 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1321 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1381 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1441 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1501 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1561 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1621 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     1681 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     1741 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     1801 ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa
     1861 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1921 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
     1981 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     2041 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     2101 atttcgcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     2161 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     2221 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     2281 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     2341 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     2401 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     2461 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     2521 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     2581 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     2641 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     2701 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     2761 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     2821 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     2881 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     2941 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     3001 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     3061 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     3121 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     3181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     3241 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     3301 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     3361 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3421 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3481 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3541 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3601 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3661 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3721 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     3781 ctgcgcagat ctgctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     3841 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     3901 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     3961 acaaacgacc caacacccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     4021 tccttccggt attgtctcct tccgtgtttc agttagcctc ccccgttcca ggcggggagg
     4081 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     4141 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     4201 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccat ggaaaggacg
     4261 tcagcttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     4321 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     4381 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     4441 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     4501 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     4561 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     4621 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgagatat caaactcatt
     4681 actaaccggt agggatcgaa cccttaagct tactagtgat gatgatgatg atgcccgcct
     4741 gtttctggca ggccgctagc atagtcaggt acatcatacg gatactttat ctcgagtttt
     4801 gttccgccgc caaaagtagg gggagagcgg tggtattgat gacaatagta tgtagcggcg
     4861 tcctctgctt ccatagaact aatcgttaag gaaaatgaag ttccgcttcc ggatccggaa
     4921 aaccgcacgg gcacgccgct ggcgagattg ctggtggaat agatccaaag cttcgggcta
     4981 gagccaggct tttgttgata ccaatgcagg taggtgctgg acacagagct ggatgctgta
     5041 caagtcattg taacgcgctc gcccagacta gctgacatta tcgccggact ttgagtcagc
     5101 acgatctgtg aaccgccgcc acttccacca ccgccggaac ctcctccgcc ggatcctcct
     5161 ccaccggaac taacagtcac ggtagttcca gctccccata cgtcaaaata gccggatgag
     5221 ctcccataat atcttctccg ggagcaatag taagtcgcgg tatcttctgt ggttacactg
     5281 ttgagttgaa gaaagaattg attcttactg gtatcacggg taattgacac tctgctcttt
     5341 aagctcggat tataagacgt actgtctgag aaactgatat aacccatcca ttcgagctta
     5401 ttccctggga attgtctaat ccaataccat gcgtagtcag acgttatact ataaccggtc
     5461 acggtacatg ttaagctgag tgattgggat gggcggacca accccgggcc agattcttgc
     5521 aattgtacgt cggaatggac tccagtcgcg gtcgctacga gaaagagaat tatacagctc
     5581 cagcccatgg tggcggcggg aattcaaggg ttcgatcctc tagagtccgg aggctggatc
     5641 ggtcccggtg tcttctatgg aggtcaaaac agcgtggatg gcgtctccag gcgatctgac
     5701 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     5761 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     5821 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     5881 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     5941 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     6001 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     6061 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     6121 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     6181 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     6241 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcaac
     6301 gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta agcagattct
     6361 tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact
     6421 cagcgacctc caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc
     6481 agattgtact gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt
//
