LOCUS       sequence-418363-        6886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression Plasmid of Ankyrin-R scFv. Derived from 
            hybridoma N388A/10..
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: Ankyrin-R scFv [N388A/10]
FEATURES             Location/Qualifiers
     source          1..6886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     597..616
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(835..939)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1014..1033)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1205..1584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1585..1788
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1739..1759
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1785..1809
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1908..1917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2694..2720
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2745..2762
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2818..3406
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2871..2891)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3289..3300)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3427..3446)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3471..3519
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3721..4149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3808..3827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4018..4039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4158..4178)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4163..4492
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4343..4478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4405..4424
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4559..5353
                     /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(4613..4632)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5223..5242
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5529..5650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5566..5585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5620..5639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5699..5715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5699..5715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5712..5734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5723..5739
                     /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(5747..5753,5754..5771,5772..5777))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5792..5813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5930..5947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6101..6689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6181..6200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6860..6886,1..765,766..834))
                     /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"
ORIGIN
        1 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
       61 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      121 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      181 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      241 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      301 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      361 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      421 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
      481 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
      541 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
      601 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
      661 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
      721 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
      781 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      961 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     1021 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     1081 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     1141 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     1201 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     1261 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1321 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     1381 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     1441 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     1501 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1561 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     1621 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     1681 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     1741 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     1801 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     1861 cgatccagcc tccggactct agaggatcga acccttgaat tcccgccgcc accatgggct
     1921 ggagctgtat tattcttttc ttggtggcta ccgctacagg ggttcattcc gaggttaaac
     1981 tcgtagaatc cggcggtggt ttggtaagac caggtggaag ccttaagctt tcatgcgccg
     2041 caagcgggtt tacattttct tcttacgcta tgagctgggt ccgtcaaacc ccggagaaaa
     2101 gattggaatg ggtggccagc atctcttcag gcggacggac ttattaccct gattctgtta
     2161 aaggtcggtt tacgattagc cgagacgacg cgagaaatat tctctatctc cagatgtcta
     2221 gcttgagatc agaagatacc gctatgtact attgcgcccg agattcttac gagttcgcat
     2281 attggggaca gggaaccctc gttacggttt ccgccggtgg cggtggttct ggtggtggtg
     2341 gatcaggggg gggtggaagt ggcggcggta gtgatataca aatgacccag tcctcaagct
     2401 atctgagcgt gagcctgggc gggcgcgtta ctatcacatg taaagccagc gatcatatca
     2461 acaactggtt ggcatggtac caacagaagc ccgggaacgc accaagactg ctcatcagtg
     2521 gcgccacaaa cctggagacg ggagtgccaa gccggttttc cggaagcggt agtggtaaag
     2581 actataccct gtccatcact tctttacaaa cagaggacgt agcaacatac tattgccagc
     2641 aatactggtc aaccccatat acatttggcg gcggaactaa attggagatt aagtatccct
     2701 atgatgttcc tgactatgcg agtggtctgc cggaaacagg tggccatcat catcatcatc
     2761 actagtaagc ttaagggttc gatccctacc ggttagtaat gagtttgata tctcgacaat
     2821 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2881 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2941 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3001 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3061 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3121 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3181 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3241 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3301 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3361 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aacgggggag
     3421 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3481 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3541 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3601 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3661 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     3721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3841 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3901 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3961 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4021 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4081 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4141 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4201 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4321 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4381 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4441 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4501 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4561 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4621 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4681 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4741 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4801 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4861 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4921 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4981 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5041 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5101 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5161 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5221 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5281 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5341 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5401 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5461 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5521 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5641 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5701 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5761 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5821 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5881 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6541 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacc
//
