LOCUS       sequence-418368-        6892 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression Plasmid of SynDIG1/Tmem90b scFv. Derived from 
            hybridoma L42/17..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  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 6892)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: SynDIG1/Tmem90b scFv [L42/17]
FEATURES             Location/Qualifiers
     source          1..6892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..120
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(129..557)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(239..260)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     451..470
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    759..807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     832..851
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     misc_feature    872..1460
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             978..989
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1387..1407
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1516..1533)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1558..1584)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2367..2376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2475..2499)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2496..2699)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2525..2545)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2700..3079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3251..3270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3345..3449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3450..3518,3519..4310)
                     /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(3668..3687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4481..5069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4970..4989
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5223..5240
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5357..5378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5393..5398,5399..5416,5417..5423)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5431..5447
                     /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     5436..5458
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5455..5471
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5455..5471
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5520..5641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5531..5550)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5585..5604
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5817..6611)
                     /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(5928..5947)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6538..6557
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6678..6892,1..115))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6692..6827
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6746..6765)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
       61 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      121 acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      181 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
      241 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
      301 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
      361 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
      421 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
      481 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      541 gccgctacag ggcgcgtggg gataccccct agagccccag ctgcgcagat ctgctatggc
      601 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
      661 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
      721 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacacccgt
      781 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
      841 tccgtgtttc agttagcctc ccccgttcca ggcggggagg cggcccaaag ggagatccga
      901 ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag caggccgcgg
      961 gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt cccgcgcaga
     1021 atccaggtgg caacacaggc gagcagccat ggaaaggacg tcagcttccc cgacaacacc
     1081 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     1141 atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag gagctgacag
     1201 gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca caccacgcca
     1261 cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga tttatacaag
     1321 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     1381 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     1441 ttgtaatcca gaggttgatt gtcgagatat caaactcatt actaaccggt agggatcgaa
     1501 cccttaagct tattagtgat gatgatgatg atgcccacca gtttctggca ggccggaagc
     1561 atagtcaggt acatcatatg gatacttaat ctccagttta gtgcctgaac caaaggtaaa
     1621 gggcacgtgg gatccctgga aacaatagta cacacctaag tcttccgctt cgactcgact
     1681 aattttcaag gtaaagtcag tgccactgcc ggagccggaa aaccgatcgg gcacacctga
     1741 gaacctatta gagaccttat aaataagcag tttaggtgat tgtccgggct tttgaagaaa
     1801 ccactcgagg tatgtattac cgttggagtg gacgatactt tgtgaactac gacaactaat
     1861 agatgcctgg tccccgagtg atactggtaa gcttaaaggg gtctgagtca ttaacacgtc
     1921 ggaccctcct ccggacccgc caccgcctga accaccaccg ccagatccac ctcctccgga
     1981 actcacggta agggttgttc cctgacccca atagttgccg ttgctgcaat agtacaccgc
     2041 gctatcttcg ctggtgagtg aactaagctg catatacgcg ctattagaag aggtgtccac
     2101 tgtaaaagta gcttttcctt taaacttttc gttataattt gtcctgccgc tgcccgggaa
     2161 gatatcccct atccattcga gtccgtgccc gggtctttgt ttgatccatt cgatccaata
     2221 atcgttaaaa gtatatcccg tcgctttaca actcatttta acggatgcgc ctggtttcat
     2281 aagttcggcg cctgattgtt gcaattgtac ttgagaatgg actccagtag cggtagccac
     2341 aagaaacaaa ataatacagc tccagcccat ggtggcggcg ggaattcaag ggttcgatcc
     2401 tctagagtcc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     2461 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac
     2521 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     2581 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     2641 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     2701 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     2761 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     2821 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     2881 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     2941 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     3001 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     3061 ctagtcaata atcaatgtca acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa
     3121 cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca agccttgcct tgttgtagct
     3181 taaattttgc tcgcgcacta ctcagcgacc tccaacacac aagcagggag cagatactgg
     3241 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat
     3301 ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3421 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3481 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3541 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3601 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3661 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3721 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3781 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3841 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3901 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3961 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4021 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4081 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4141 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4201 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4261 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4321 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4381 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4441 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4501 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4561 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4621 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4681 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4741 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4801 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4861 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4921 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4981 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5041 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5101 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5161 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5221 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5281 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5341 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5401 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5461 acagctatga ccatgattac gccaagctct agctagaggt cgacggtata cagacatgat
     5521 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5581 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5641 tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     5701 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     5761 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgcg aaccccagag tcccgctcag
     5821 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     5881 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     5941 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     6001 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     6061 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     6121 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     6181 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     6241 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     6301 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     6361 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     6421 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     6481 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     6541 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     6601 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatccgaa aatggatata
     6661 caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     6721 tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     6781 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     6841 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gc
//
