LOCUS       sequence-397521-        4912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tagging endogenously expressed proteins with ""spaghetti monster"" 
            smFP-V5, C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4912)
  AUTHORS   Gao Y, Shonai D, Trn M, Zhao J, Soderblom EJ, Garcia-Moreno SA, 
            Gersbach CA, Wetsel WC, Dawson G, Velmeshev D, Jiang YH, Sloofman 
            LG, Buxbaum JD, Soderling SH
  TITLE     Proximity analysis of native proteomes reveals phenotypic modifiers 
            in a mouse model of autism and related neurodevelopmental 
            conditions.
  JOURNAL   Nat Commun. 2024 Aug 9;15(1):6801. doi: 10.1038/s41467-024-51037-x.
  PUBMED    39122707
REFERENCE   2  (bases 1 to 4912)
  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: "Nat 
            Commun."; date: "2024-08-9"; volume: "15(1):6801. doi"; pages: " 
            10.1038/s41467-024-51037-x"
COMMENT     Sequence Label: pHiUGE smFP-V5 donor (all-in-one) ORF0
FEATURES             Location/Qualifiers
     source          1..4912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..393
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        414..489
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        501..749
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        768..843
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(888..929)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(939..980)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(990..1031)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1245..1286)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1296..1337)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1356..1397)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1407..1448)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1980..2021)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2031..2072)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2082..2123)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     repeat_region   2175..2315
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2175..2304
                     /label=AAV2 ITR
     rep_origin      2390..2845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2477..2496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2687..2708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2862..2881)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2981..3003
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3041..3059)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3127..3231
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3232..3300,3301..4092)
                     /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(3450..3469)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4263..4851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4752..4771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat
      181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      361 gctttatata tcttgtggaa aggacgaaac accgacgctt ccgagtacgg tacgttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      481 agtcggtgct ttttttctag gagggcctat ttcccatgat tccttcatat ttgcatatac
      541 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
      601 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      661 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      721 tatatatctt gtggaaagga cgaaacaccg gaagagcgag ctcttctgtt ttagagctag
      781 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
      841 tgcttttttt ctagacctgt accgtactcg gaagcgtcgg atccttaggt actatccagt
      901 cccagcaacg ggttcgggat gggcttgccc tgctgttgcg tactgtcgag gcccaacagt
      961 ggatttggta ttggctttcc ttgttgctga gtgctgtcga ggcccagcaa gggatttggt
     1021 atgggcttgc ctccttggcc cttgtacagc tcgtccattc ccaacgtaat gccggcggcg
     1081 gtcacgaact ccaacaacac catgtgatct cgtttttcgt ttgggtcctt actcagaacg
     1141 ctctgagtcg acagatagtg attatccggc aacagcacgg ggccatctcc gatcggcgta
     1201 ttctgctgat agtggtccgc cagttgaaca cttccatcgc cacctgtcga gtcgaggccc
     1261 agcagagggt tggggattgg cttgccttgc tgctgggtgc tatccaggcc gagcagggga
     1321 ttgggtatcg gtttgccacc gccttgctgg cctccggtgc tgtcgagtcc cagcaagggg
     1381 ttagggatgg gctttccctg ttgctgcgtg ctatccaagc ccagcagggg attcgggatc
     1441 ggcttgccac caccttccac gttgtgccga attttgaaat tcgcctttat gccgttcttc
     1501 tgcttgtcgg cagtaatgta gacgttatgg ctgttaaagt tgtattccag cttgtgaccc
     1561 agaatattac cgtcttcctt gaaatcaatt cccttaagtt ctatcctgtt caccaaggtg
     1621 tctccctcga acttcacctc ggctcgcgtc ttgtaagtac cgtcatcttt gaaactgatc
     1681 gtccgttcct gcacgtatcc ctccggcata gcactcttaa agaagtcatg ctgcttcatg
     1741 tggtctgggt aacggctgaa acactggact ccgccaccca gggttgtaac caaggtcggc
     1801 cacggcaccg gcagctttcc cgtggtgcat ataaacttca gcgtcaattt tccgttggtg
     1861 gcatctccct ctccttctcc gcgcacacta aacttgtgtc cgttaacgtc accgtcgagc
     1921 tcaaccagaa ttgggaccac gccggtaaac agttcttcgc ccttactcac accttggccc
     1981 gtggagtcca ggcccaacag agggttaggg ataggcttac cttgttgctg tgtgctatcc
     2041 agaccgagaa gtgggtttgg aatgggctta ccctgctgct gagtactatc aagtccaagg
     2101 agggggttcg gtataggttt tccctcgagc ctgtaccgta ctcggaagcg tccacgtgcg
     2161 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2221 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2281 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2341 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2401 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2461 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2521 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2581 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2641 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2701 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2761 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2821 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     2881 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2941 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3001 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3061 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3121 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3181 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3241 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3301 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3361 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3421 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3481 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3541 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3601 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3661 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3721 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3781 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3841 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3901 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3961 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4021 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4081 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4141 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4261 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4321 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4381 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4441 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4501 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4561 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4621 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4681 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4741 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4801 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4861 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
