LOCUS       sequence-439004-        7170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Sun1GFP for nuclei isolation and Projection-TAG BC for
            multiplex projection tracing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7170)
  AUTHORS   Yang L, Liu F, Hahm H, Okuda T, Li X, Zhang Y, Kalyanaraman V, 
            Heitmeier MR, Samineni VK
  TITLE     Projection-TAGs enable multiplex projection tracing and multi-modal 
            profiling of projection neurons.
  JOURNAL   Nat Commun. 2025 Jul 1;16(1):5557. doi: 10.1038/s41467-025-60360-w.
  PUBMED    40593543
REFERENCE   2  (bases 1 to 7170)
  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: "2025-07-1"; volume: "16(1):5557. doi"; pages: " 
            10.1038/s41467-025-60360-w"
COMMENT     Sequence Label: pAV-CAG-Sun1-GFP-WPRE-BC1-pA
FEATURES             Location/Qualifiers
     source          1..7170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     1001..1023
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1085..1104
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(2802..2804,2805..3518)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2802..3515
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2832..2860)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3455..3475
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    3536..4124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3589..3609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4007..4018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4267..4388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4278..4297)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4332..4351
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4894..5482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4974..4993)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5653..6444,6445..6513))
                     /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     6276..6295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6514..6618)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6644..7099)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6781..6802)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6993..7012
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg gggcacccca ggcggggggg ggcggggcgg ggcgaggggc
      601 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      661 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      721 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      781 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      841 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      901 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg
      961 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1021 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1081 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccgaat
     1141 tcactagcat gacctccagg gtgtactcca gagacaggac tctcaaacca cggaaggcag
     1201 cctcgggaac cttctggtgg ctagggagcg gctggtacca atttgttact ttgatttctt
     1261 ggctgaatgt ctttcttctt accaggtgcc ttcgaaatat ttgcaaggtt tttgtcttgc
     1321 tcctcccact cctactttta ctaggtgctg gtgtctccct gtggggccag ggaaacttct
     1381 tctcactcct accagtgctg aactggacgg ccatgcagcc aacacagagg gtggacgatt
     1441 ccaagggcat gcatagacct ggccctcttc ccccgagccc acctccaaag gttgatcaca
     1501 aggcttccca gtggcctcag gagagtgaca tggggcagaa ggtagcttct ttgagtgcgc
     1561 agtgccacaa ccatgatgag agacttgcag agctgacagt cctgcttcag aaactacaga
     1621 tacgggtaga ccaagtggat gacggcaggg aagggctgtc actgtgggtc aagaatgtgg
     1681 ttggacagca cctgcaggag atgggcacca tagaaccacc tgatgctaag actgacttca
     1741 tgactttcca ccatgaccat gaagtgcgtc tctccaactt ggaagatgtt cttagaaaac
     1801 tgacagaaaa atctgaggct atccagaagg agctggaaga aaccaagctg aaagcaggca
     1861 gcagggatga agagcagccc ctccttgacc gtgtgcagca cctagaactg gaactgaacc
     1921 tgttgaagtc acagctgtca gactggcagc atctgaagac cagctgtgag caggctgggg
     1981 cccgcatcca ggagactgtg cagctcatgt tctctgagga tcagcagggc ggttccctcg
     2041 agtggctatt agagaagctt tcttctcggt tcgtgagcaa ggatgagctg caggtgctct
     2101 tacatgacct tgagctgaaa ctgctgcaga atatcacaca ccacatcacc gtgacaggac
     2161 aggccccgac atccgaggct attgtgtctg ccgtgaatca ggcagggatt tcaggaatca
     2221 cagaagcgca agcacatatc attgtgaaca atgctctgaa gctgtactcc caagacaaga
     2281 cggggatggt ggactttgct ctggagtctg gaggtggcag catcctaagc actcggtgct
     2341 ctgagaccta tgagaccaag acggcactgc tgagcctgtt tggggtccca ctgtggtact
     2401 tctcacagtc acctcgagtg gtgatccagc ccgacatcta cccagggaat tgctgggcgt
     2461 tcaaaggttc ccaggggtac ctggtggtgc ggttgtccat gaagatctac ccaaccacat
     2521 tcaccatgga acacattcca aagacactat cacccactgg taacatctcc agtgccccca
     2581 aagactttgc agtctatgga ctggaaacgg agtatcaaga agaggggcag cctctgggac
     2641 ggttcaccta tgaccaggaa ggagactcac tccagatgtt ccacacactg gaaagacctg
     2701 accaagcctt ccagatagta gagctccggg tcctgtccaa ctggggccac cctgagtaca
     2761 cttgcctcta ccggttccga gtccacggag agcccatcca gatgagcaaa ggagaagaac
     2821 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
     2881 tttctgtccg tggagagggt gaaggtgatg ctacaaacgg aaaactcacc cttaaattta
     2941 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ctgacctatg
     3001 gtgttcaatg cttttcccgt tatccggatc acatgaaacg gcatgacttt ttcaagagtg
     3061 ccatgcccga aggttatgta caggaacgca ctatatcttt caaagatgac gggacctaca
     3121 agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaaag
     3181 gtattgattt taaagaagat ggaaacattc tcggacacaa actcgagtac aactttaact
     3241 cacacaatgt atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa
     3301 ttcgccacaa cgttgaagat ggttccgttc aactagcaga ccattatcaa caaaatactc
     3361 caattggcga tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctgtcc
     3421 tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg
     3481 ctgggattac acatggcatg gatgagctct acaaataagg cgcgagctta tcgataatca
     3541 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3601 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3661 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3721 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3781 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3841 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3901 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     3961 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4021 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4081 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     4141 cgggcggccg cacctgtaga acaaccaggt gatgttccac gtggacatct agcaggtgct
     4201 gatcgagaag gactacttga acgacatgct ctagctctag atctacgtct tttcgagcag
     4261 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4321 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4381 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     4441 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     4501 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     4561 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgcccgggct
     4621 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg
     4681 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4741 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4801 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4861 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4921 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4981 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5041 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5101 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5161 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5221 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5281 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5341 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5401 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5461 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5521 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5581 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5641 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5701 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5761 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5821 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5881 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5941 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6001 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6061 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6121 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6181 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6241 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6301 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6361 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6421 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6481 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6541 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6601 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     6661 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6721 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6781 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6841 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6901 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6961 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7021 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7081 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7141 tcggtgcggg cctcttcgct attacgccag
//
