LOCUS       sequence-439274-        5485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of oScarlet for cell 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 5485)
  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 5485)
  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-oScarlet-WPRE-BC11-pA
FEATURES             Location/Qualifiers
     source          1..5485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     995..1017
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1079..1098
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1137..1832
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWDRVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1270..1288)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1538..1557
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1865..2453
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1918..1938)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2336..2347)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2577..2698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2588..2607)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2642..2661
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3209..3797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3289..3308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3968..4759,4760..4828))
                     /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     4591..4610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4829..4933)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4959..5414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5096..5117)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5308..5327
                     /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 ggcgcgccrg gggggggggg gggggggggg ggggggggsg
      601 gggsgrggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      661 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      721 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      781 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      841 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      901 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
      961 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1021 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1081 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc gaattcatgg
     1141 tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct
     1201 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1261 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc tgggacatcc
     1321 tgtcccctca gttcatgtac ggctccaggg ccttcaccaa gcaccccgcc gacatccccg
     1381 actactataa gcagtccttc cccgagggct tcaagtggga ccgcgtgatg aacttcgagg
     1441 acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc ctgatctaca
     1501 aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa
     1561 tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg aagggcgaca
     1621 ttaagatggc cctgcgcctg aaggacggcg ggcgctacct ggcggacttc aagaccacct
     1681 acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtggaccgc aagttggaca
     1741 tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc gagggccgcc
     1801 actccaccgg cggcatggac gagctgtaca agtaaaagct ttctagaggc gcgagcttat
     1861 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2041 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2101 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2161 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2221 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2281 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2341 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2401 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgcggccg
     2461 cagaacttgc accacgacat cgagctccag gtcgtcgtca tcatcgtcat ggaggacttg
     2521 gtcgacgacg tctagtaggt ggacgagaag aagcagttgg tttcgagcag acatgataag
     2581 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2641 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2701 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     2761 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     2821 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     2881 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgnnnnnccc gggcaaagcc
     2941 cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg
     3001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3961 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4021 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4081 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4141 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4201 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4261 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4321 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4381 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4441 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4501 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4561 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4621 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4681 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4741 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4801 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4861 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4921 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     4981 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5041 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5101 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5161 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5221 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5281 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5341 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5401 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5461 gcgggcctct tcgctattac gccag
//
