LOCUS       sequence-418281-        6925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of QKI-6 scFv. Derived from hybridoma 
            N182/17..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  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 6925)
  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: QKI-6 scFv [N182/17]
FEATURES             Location/Qualifiers
     source          1..6925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      439..448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(547..571)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(568..771)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(597..617)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        772..1151
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1323..1342
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1417..1521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1522..1590,1591..2382)
                     /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(1740..1759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2553..3141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3042..3061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3295..3312
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3429..3450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3465..3470,3471..3488,3489..3495)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3503..3519
                     /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     3508..3530
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3527..3543
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3527..3543
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3592..3713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3603..3622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3657..3676
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3889..4683)
                     /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(4000..4019)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4610..4629
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4750..5079)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4764..4899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4818..4837)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5064..5084
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5093..5521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5203..5224)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5415..5434
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5723..5771
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5796..5815
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     misc_feature    5836..6424
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5942..5953
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6351..6371
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6480..6497)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(6522..6548)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 cactgccgcc gccgcctgca gagacagtga ccagagtccc ttggccccag taagcaaacc
       61 aggtacccca tagaagatcc ccttctcttg cacagaaata agtggcactg tcctcagttc
      121 tcagggtgtt cagttgaaga tagaggatgc tttgggaatt atctctggag atggtgaacc
      181 gacccctcac agatgcactg tattctgttg tgtaaccatt agctttgttt ctagtaaaac
      241 ccaaccactc aagtgccttt cctggaggct ggcggaccca actcatgtag taatcactga
      301 aggtgaaccc agaagttgca caggagagtc tcagagaacc cccaggctgt tccaagcctc
      361 ctccagactc caccagggtc acctcgctgt gcacgccggt ggctgtggcg accaggaaca
      421 ggatgataca gctccagccc atggtggcgg cgggaattca agggttcgat cctctagagt
      481 ccggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      541 ccaggcgatc tgacggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
      601 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
      661 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
      721 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
      781 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
      841 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
      901 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
      961 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     1021 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     1081 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     1141 taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac
     1201 cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt
     1261 gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta
     1321 tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat
     1381 ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1441 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1501 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1621 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1681 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1741 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1801 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1921 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2041 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2101 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2221 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2341 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3241 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3301 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3361 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3421 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     3481 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     3541 gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca
     3601 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3661 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg
     3721 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     3781 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
     3841 aggttgggcg tcgcttggtc ggtcatttcg cgaaccccag agtcccgctc agaagaactc
     3901 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     3961 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     4021 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     4081 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     4141 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     4201 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     4261 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     4321 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     4381 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     4441 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     4501 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     4561 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     4621 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     4681 catgcgaaac gatcctcatc ctgtctcttg atcagatccg aaaatggata tacaagctcc
     4741 cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     4801 ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     4861 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     4921 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4981 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5041 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg
     5101 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5161 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5221 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5281 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5341 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5401 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5461 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5521 tggggatacc ccctagagcc ccagctgcgc agatctgcta tggcagggcc tgccgccccg
     5581 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     5641 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     5701 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac ccgtgcgttt tattctgtct
     5761 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     5821 cctcccccgt tccaggcggg gaggcggccc aaagggagat ccgactcgtc tgagggcgaa
     5881 ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc gcgggaagga aggtccgctg
     5941 gattgagggc cgaagggacg tagcagaagg acgtcccgcg cagaatccag gtggcaacac
     6001 aggcgagcag ccatggaaag gacgtcagct tccccgacaa caccacggaa ttgtcagtgc
     6061 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
     6121 caatagggag ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc
     6181 aaccagtggg ggttgcgtca gcaaacacag tgcacaccac gccacgttgc ctgacaacgg
     6241 gccacaactc ctcataaaga gacagcaacc aggatttata caaggaggag aaaatgaaag
     6301 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     6361 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     6421 gattgtcgag atatcaaact cattactaac cggtagggat cgaaccctta agcttatcaa
     6481 tggtggtggt ggtggtgtcc gccggtctca gggagcccag aggcgtaatc tggcacgtcg
     6541 taagggtatt ttatttccaa ctttgtcccc gagccgaacg tgaatggaac atgtgaacct
     6601 tgaaagcagt aataaactcc cagatcctca gcctccactc tgctgatctt gagtgtgaaa
     6661 tctgtccctg atccactgcc actgaacctg tctgggaccc cagaaaatcg gttggaaact
     6721 ctgtagatca ggagctttgg agactggcct ggtttctgca ggtaccattc taaaaaggtg
     6781 attccaatac tatgtacaat ggtctgactc gatctgcaag agatggaggc ttgatctcca
     6841 agactgacag gcagggagag tggagtttgg gtcatcaaaa catcagagcc gcctccagag
     6901 cctcctccgc cgctgcctcc gcctc
//
