LOCUS       sequence-438061-        7131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-STARR-seq validation vector containing VS33 enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7131)
  AUTHORS   Chan YC, Kienle E, Oti M, Di Liddo A, Mendez-Lago M, Aschauer DF, 
            Peter M, Pagani M, Arnold C, Vonderheit A, Schon C, Kreuz S, Stark 
            A, Rumpel S
  TITLE     An unbiased AAV-STARR-seq screen revealing the enhancer activity map
            of genomic regions in the mouse brain in vivo.
  JOURNAL   Sci Rep. 2023 Apr 25;13(1):6745. doi: 10.1038/s41598-023-33448-w.
  PUBMED    37185990
REFERENCE   2  (bases 1 to 7131)
  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: "Sci Rep."; 
            date: "2023-04-25"; volume: "13(1):6745. doi"; pages: " 
            10.1038/s41598-023-33448-w"
COMMENT     Sequence Label: 
            pAAV_Seq17_VS33_4.26_ChimeraIntron_3NLS_RFP_WPRE_polyA
FEATURES             Location/Qualifiers
     source          1..7131
                     /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        2209..2239
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          2266..2398
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2381..2400
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2437..2457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2458..2478
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2485..2505
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2506..3219
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    3256..3844
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3309..3329)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3727..3738)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3847..3863)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3848..3864)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3878..4354
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4394..4534
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4394..4523
                     /label=AAV2 ITR
     rep_origin      4609..5064
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4696..4715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4906..4927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5081..5100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5200..5222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5260..5278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5346..5450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5451..5519,5520..6311)
                     /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(5669..5688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6482..7070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6971..6990
                     /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 gcgtagataa tgagggtcag ttcagaggcc attgcacgga
      181 ccatgacccc ttgtcaagag agcagagcca atctactatt atttctgcta aatggacatg
      241 gtgcatgggt tagtgcatca aacaagcccc atcaaggaag cttctttttt gaagcagata
      301 gcatgtaacg cagagaacca caactggcca tggtgcagag aataagagac tacagaatac
      361 tcagccctag atgtaatgtc tgtctcacac tcctcccgtc aaggctgaga gaactttgaa
      421 gaaaaggggg cagcaagaat caatggggat gacattaact gtgaaactgt gattcacaac
      481 attgtggata tggaacctga agaggctatc tcctgtaccc cagtgcagtg atacagacac
      541 caacccagcc acaaaacttt caacccaaaa tttatcctgt ctacacgaag tgtaggcacg
      601 gggcagggag cagagactga cggaacgcgt gagcgttcct ttctcagctc cttggccagg
      661 caggcgctcg cggctacctg ccctgcttgg cgccgcactg aagattcgcc gcggcggaac
      721 agttgagtct ccatctggct accaacccac ccgagctttc ttctcctcca tcaccacctt
      781 cctgcctccc cctcctcccc ctcctttccg tcttccctct ccacccccgc ccccaatctc
      841 ctcctctttt tctcactaca agcggttgct gatgctgaag ccgagcgtca cttcggctcc
      901 cacggcagac atggcgacat tgacagtggt ccagccgctt actctggaca gaggtaaggg
      961 agcggctggt ccgtcaggcc acacagtccg caccgcttcg ccctccgccc tgcccagtcc
     1021 cacagggaag gcaggatgcg ctcgggcagg ggcagctggg ggaggaggat cagcccagga
     1081 gggtttcgcg cccggagcag gacactttct tcccctctct cctctcctgg tccccgctgc
     1141 cccttcttgc tgcgctctcc ggggcgtagg tgcttgccag ggctcgaatg tttcagctga
     1201 cagtctcggt ccctcgccgg gtcgcctccg ggccccggct ttgttggaga ggatgcgccg
     1261 ggaccgctga gcgctggtga ctggaggttg aagaggggca gagaaagctc tcccagccag
     1321 agcttgctaa gggaccagtg gggaagctag ctgccaactg ccggcgcggg gacccgaagg
     1381 gaggaagttt gtccccgcgc tggagatgag gggtctgtgc agctgttgtg actgggagat
     1441 cgccccaggg cggcagggcg cttggctggg aagctgggtg caaagccgcg ggactgcctg
     1501 gctagggcgc tgccctcggg cgaaagggca gagcagagct cgggcaacct tcgctggctc
     1561 agggacaccg catccggagc cgagccggct tggctccagc ttcccgctcg gagctgcggc
     1621 tctggtcttg gtactccctc agcctgcagt tccagggagg tagcaactcc tgtctcctgc
     1681 tccagcaaac cttgttcttc accagtcaag tgtacctcag tgcctgggct gcagaaggcg
     1741 cggagacaaa gcgcaggctg ggtaactgtt taacttctag cctgtgacct gtgcgtccta
     1801 ctctttggct gaaacggaaa gagcttttgc tcaaaccttt ccaaaccttt ccaaaccttt
     1861 ctctaggatc gggccctgct gagtgcaccc gagtagtacc tggcggtgga tgggagtttc
     1921 ctttctgcct caccaggtcc agttagttat tagggctcca gagtccccag ctctgcagag
     1981 agagggcacc tctagctctg ggtccgcagc ggcggggctt aggacccggt gcagtcattt
     2041 gaaaatgctg tgctcagaag aaaatgattt agtccttgtc cctctaaaat ttctctttct
     2101 ggaggattaa actagcggcc cagcaaagca aaaagagcat cgtacagcga gcacagtggc
     2161 aaagctggta aactgtattt gaggttctta atgtggaacc ggttcgagag agggtatata
     2221 atggaagctc gacttccagg ggcccggtcg tgaggcactg ggcaggtaag tatcaaggtt
     2281 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     2341 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     2401 gtccaccctg caggcttaag catcgatacc atggacccaa aaaagaagcg gaaggtccca
     2461 aaaaagaagc ggaaggtcga attcccaaaa aagaagcgga aggtcatggt gtctaagggc
     2521 gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtgaacaac
     2581 caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
     2641 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc
     2701 ttcatgtacg gcagcagaac cttcatcaac cacacccagg gcatccccga cttctttaag
     2761 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
     2821 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc
     2881 agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact cggctgggag
     2941 gccaacaccg agatgctgta ccccgctgac ggcggcctgg aaggcagaag cgacatggcc
     3001 ctgaagctcg tgggcggggg ccacctgatc tgcaacttca agaccacata cagatccaag
     3061 aaacccgcta agaacctcaa gatgcccggc gtctactatg tggaccacag actggaaaga
     3121 atcaaggagg ccgacaaaga gacctacgtc gagcagcacg aggtggctgt ggccagatac
     3181 tgcgacctcc ctagcaaact ggggcacaaa cttaattgag gatcctctag agtcgacctg
     3241 cagaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3301 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3361 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3421 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3481 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3541 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3601 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3661 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3721 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3781 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3841 ccgcatcgat accgtcgacc tcgagaccta gatctacggg tggcatccct gtgacccctc
     3901 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3961 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     4021 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     4081 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     4141 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     4201 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     4261 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     4321 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     4381 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4441 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4501 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4561 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4621 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4681 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4741 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4801 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4861 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4921 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4981 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5041 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5101 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5161 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5221 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5281 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5341 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5401 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6001 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6061 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6121 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6181 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6241 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6301 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6361 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6421 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6481 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6541 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6601 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6661 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6721 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6781 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6841 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6901 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6961 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7021 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7081 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
