LOCUS       sequence-397390-        4913 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 4913)
  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 4913)
  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) ORF2
FEATURES             Location/Qualifiers
     source          1..4913
                     /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   2176..2316
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2176..2305
                     /label=AAV2 ITR
     rep_origin      2391..2846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2478..2497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2688..2709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2863..2882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2982..3004
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3042..3060)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3128..3232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3233..3301,3302..4093)
                     /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(3451..3470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4264..4852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4753..4772
                     /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 tcccctcgag cctgtaccgt actcggaagc gtccacgtgc
     2161 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2221 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2281 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2341 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2401 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2461 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2521 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2581 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2641 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2701 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     2761 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2821 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2881 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2941 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3001 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3061 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3121 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3181 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3241 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3301 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3361 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3421 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3481 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3541 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3601 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3661 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3721 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3781 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3841 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3901 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3961 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4021 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4081 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4141 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4381 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
