LOCUS       sequence-397525-        4914 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 4914)
  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 4914)
  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) ORF1
FEATURES             Location/Qualifiers
     source          1..4914
                     /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   2177..2317
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2177..2306
                     /label=AAV2 ITR
     rep_origin      2392..2847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2479..2498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2689..2710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2864..2883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2983..3005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3043..3061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3129..3233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3234..3302,3303..4094)
                     /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(3452..3471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4265..4853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4754..4773
                     /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 tccctctcga gcctgtaccg tactcggaag cgtccacgtg
     2161 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2221 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2281 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2341 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2401 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2461 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2521 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2581 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2641 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2701 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2761 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2821 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2881 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2941 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3001 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3061 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3121 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3181 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3421 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4081 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4141 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4201 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4261 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4321 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4381 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4441 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4501 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4561 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4621 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4681 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4741 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4801 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4861 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
