LOCUS       sequence-418367-        6886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of Mitofusin-1 scFv. Derived from 
            hybridoma N111/24..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6886)
  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 6886)
  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: Mitofusin-1 scFv [N111/24]
FEATURES             Location/Qualifiers
     source          1..6886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(11..802,803..871))
                     /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     634..653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(872..976)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1051..1070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1242..1621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1622..1825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1776..1796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1822..1846
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1945..1954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2731..2757
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2782..2799
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2855..3443
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2908..2928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3326..3337)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3464..3483)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3508..3556
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3758..4186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3845..3864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4055..4076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4195..4215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4200..4529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4380..4515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4442..4461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4596..5390
                     /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(4650..4669)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5260..5279
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5566..5687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5603..5622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5657..5676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5736..5752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5736..5752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5749..5771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5760..5776
                     /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(5784..5790,5791..5808,5809..5814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5829..5850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5967..5984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6138..6726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6218..6237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
       61 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      121 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      181 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      241 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      301 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
      361 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      421 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      481 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      541 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      601 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
      661 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      721 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
      781 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
      841 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      901 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      961 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     1021 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     1081 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     1141 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     1201 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     1261 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     1321 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     1381 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     1441 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1501 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1561 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1621 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1681 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1741 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1801 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     1861 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctaga
     1921 ggatcgaacc cttgaattcc cgccgccacc atgggctgga gctgtataat tttgttcttg
     1981 gtggcgaccg caacaggagt acattcagag gttaagctcg aagaaagtgg cggcgggctg
     2041 gtccagccag gccgcagcat gaagttgtcc tgcgcagcgt ccggctttag cttctccgat
     2101 gcttggatgg attgggtgag gcaatccccc gaaaagggtc tggaatgggt ggccgagatc
     2161 aggtctaagg cgaacaacca cgctacttat tacgcggaat ccgtaaaggg cagatttact
     2221 atatcccgtg acgacagcaa gtcttccgtg tatttacaga tgaattctct tagggcagag
     2281 gatacaggaa tatactattg caccgtggtg gctggaaatt attggggtca agggacgaca
     2341 ttgactgtga gttccggagg aggtggatcc ggtggcggcg ggtctggtgg tggtgggtca
     2401 ggtggtggaa gccagatcat actgactcaa agtcctgcga ttatgagcgc tagtcctggc
     2461 gaaaaggtga ctattacatg ttctgcatct agcagcgtgt cttttatgta ttggtttcaa
     2521 cagaaacccg ggaccagccc gaagttttgg atctacagca cctctaatct tgccagcggg
     2581 gtgccggcca gattttccgg aagcgggagc ggcactagct atagcctgac catatcacgc
     2641 atggaagctg aggacgcagc tacctactat tgtcaacaga gaaactcata tccgtttacc
     2701 tttgggtctg gtaccaaatt agaggttaag tatccctatg atgtccccga ctatgcatcc
     2761 ggcctgcccg aaacaggcgg ccatcatcat catcatcact agtaagctta agggttcgat
     2821 ccctaccggt tagtaatgag tttgatatct cgacaatcaa cctctggatt acaaaatttg
     2881 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2941 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3001 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3061 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3121 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3181 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3241 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3301 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3361 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3421 ctccctttgg gccgcctccc cgcctggaac gggggaggct aactgaaaca cggaaggaga
     3481 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     3541 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3601 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     3661 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     3721 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3781 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3841 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3901 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3961 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4021 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4081 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4141 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4201 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4261 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4321 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4381 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4441 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4501 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4561 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4621 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4681 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4741 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4801 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4861 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4921 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4981 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5041 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5101 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5161 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5221 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5281 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5341 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5401 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5461 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5521 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5581 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5641 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5701 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5761 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5821 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5881 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5941 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6001 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6061 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6121 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6181 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6241 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6301 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6361 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6421 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6481 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6541 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6601 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6661 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6721 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6781 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6841 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaactt
//
