LOCUS       sequence-418364-        6898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of Synaptotagmin-7 scFv. Derived from 
            hybridoma N275/14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6898)
  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 6898)
  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: Synaptotagmin-7 scFv [N275/14]
FEATURES             Location/Qualifiers
     source          1..6898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             238..264
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             289..306
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    362..950
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(415..435)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(833..844)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(971..990)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1015..1063
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1265..1693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1352..1371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1562..1583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1702..1722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1707..2036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1887..2022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1949..1968
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2103..2897
                     /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(2157..2176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2767..2786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3073..3194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3110..3129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3164..3183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3243..3259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3243..3259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3256..3278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3267..3283
                     /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(3291..3297,3298..3315,3316..3321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3336..3357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3474..3491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3645..4233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3725..3744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4404..5195,5196..5264))
                     /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     5027..5046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5265..5369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5444..5463)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5635..6014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6015..6218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6169..6189
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6215..6239
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      6338..6347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atctcacact acctgagttg gtaccaacaa aagcccgacg gtacagtcaa gctgttaatt
       61 cactatacgt ctaagctgca ttctggcgtt cctagccgct ttagcgggtc aggcagtggt
      121 accgactaca gcctgactat ttcttattta gagccagagg acatagcaac gtattactgc
      181 caacagtact ctaatttgcc atatacattt ggcggcggta cgaaactcga gattaagtat
      241 ccctatgatg ttcctgacta tgctagtgga ctgccagaaa ctggtggcca tcatcatcat
      301 catcactagt aagcttaagg gttcgatccc taccggttag taatgagttt gatatctcga
      361 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
      421 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
      481 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
      541 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
      601 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
      661 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
      721 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
      781 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
      841 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
      901 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaacggg
      961 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1021 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     1081 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     1141 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     1201 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     1261 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1321 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1381 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1441 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1501 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1561 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1621 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1681 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1741 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1861 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1921 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     1981 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2041 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2101 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2161 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2221 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2281 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2341 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2401 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2461 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2521 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2581 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2641 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2701 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2761 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2821 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2881 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     2941 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3001 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3061 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3121 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3181 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     3241 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3301 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3361 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3481 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3541 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4201 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4261 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4321 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4381 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4441 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4501 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4561 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4621 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4681 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4741 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4801 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4861 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4921 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4981 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5041 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5101 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5161 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5221 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5281 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5341 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5401 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     5461 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     5521 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     5581 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     5641 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     5701 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     5761 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     5821 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     5881 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     5941 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6001 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     6061 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     6121 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     6181 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     6241 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     6301 tccggactct agaggatcga acccttgaat tcccgccgcc accatgggct ggagctgtat
     6361 tattcttttc cttgtggcta ccgcaacagg agttcattcc gaagtccaat tacaagaaag
     6421 tggccccaat ctggtcaagc ccacccaaac cctttctctg acatgctccg tgaccggtga
     6481 tagcattaca tctgactact ggaattggat aagaaagttt ccgggtaaca agttcgaata
     6541 tatgggttat atctcatatg gcggcataac ctattataac cccagtctga agagcagaat
     6601 ttcaattacg cgggatacca gtaagaatca ctactacctt cacctgaact cagtaaccac
     6661 cgaagatacg gcgacgtact attgcgtccg cgatgggtat tcaggcacaa attatgaagc
     6721 ctattgggga cagggcacat tggtgaccgt ttccgctggt ggtggcggat caggcggtgg
     6781 cggatccggt ggtggcggat ctggtggcgg gagtgacata caaatgaccc aaacgactag
     6841 ctctttgtcc gcaagcctcg gggataagat tactattagc tgttccgcgt ctcaagat
//
